Method and associated compounds for forming nanotubes

ABSTRACT

A method and associated compounds for forming nanotubes are disclosed.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation-of U.S. patent application Ser. No.10/050,292, filed Jan. 16, 2002 which claims priority to U.S.Provisional Application Ser. No. 60/262,385, filed Jan. 17, 2001.

BACKGROUND OF THE INVENTION

The present invention generally relates to a method and associatedcompounds for forming nanotubes. The present invention particularlyrelates to a method and associated compounds for the hierarchicalself-assembly of organic nanotubes from self-assembled supermacrocycles.

Nanotubes can be thought of as long, thin cylinders of carbon which areunique for their size, shape, and physical properties. For example,nanotubes are extremely strong, low weight, stabile, flexible, have goodheat conductance, and have a large surface area. In addition, nanotubespossess a host of intriguing electronic properties. The aforementionedproperties of nanotubes has lead to an intense investigation ofutilizing these structures in the fields of material science,nanotechnology, molecular electronic and photonic devices, sensor andartificial channel systems.

The particular the properties a nanotube possesses is dependent uponseveral factors including its diameter, length, chemical make up, andchirality. Therefore, the ability to produce nanotubes which haveparticular desirable properties is dependent upon being able to form orsynthesize nanotubes which have, for example a certain length.Unfortunately, prior to the present invention, the synthetic schemes forconveniently and efficiently producing nanotubes possessing a wide rangeof specific desirable characteristics have been extremely limited.

Therefore, in light of the above discussion, it is apparent that what isneeded is a method an associated compounds for forming nanotubes thataddresses the above discussed drawback of nanotube synthetic schemes.

SUMMARY OF THE INVENTION

In accordance with one embodiment of the present invention, there isprovided a compound having the formula:

wherein X is carbon or nitrogen; n is an integer of, 1, 2, 3, or 4; Y isan amino acid having an amino group covalently bound to an α-carbon ofthe amino acid and the amino group is covalently bound to a carbon ofthe (CH₂)_(n) group; and R₁ is aliphatic; and salts thereof.

In accordance with another embodiment of the present invention, there isprovided a compound having the formula:

wherein X is carbon or nitrogen; n is an integer of 1, 2, 3, or 4; R₁ isaliphatic; R₂ is hydrogen; and R₃ is —CH₃, —CH₂OH, —CH₂CH₂SCH₃,—CH₂CO₂H, —CH₂CH(CH₃)₂, —CH₂C₆H₅, —CH₂-p-(OH)C₆H₄, —CH₂CH₂CH₂CH₂NH₂, orR₂ and R₃ together form

and salts thereof.

In accordance with yet another embodiment of the present invention,there is provided a method of forming a nanotube. The method includesdisposing a compound having the formula

or salts thereof in a solution at a sufficient concentration so that thenanotube is formed, wherein X is carbon or nitrogen; n is an integer of,1, 2, 3, or 4; Y is an amino acid having an amino group covalently boundto an α-carbon of the amino acid and the amino group is covalently boundto a carbon of the (CH₂)_(n) group; and R₁ is aliphatic.

In accordance with still another embodiment of the present invention,there is provided a method of forming a nanotube. The method includesdisposing a compound having the formula

or salts thereof in a solution at a sufficient concentration so that thenanotube is formed, wherein X is carbon or nitrogen; n is an integer of1, 2, 3, or 4; R₁ is aliphatic; R₂ is hydrogen; and R₃ is —CH₃, —CH₂OH,—CH₂CH₂SCH₃, —CH₂CO₂H, —CH₂CH(CH₃)₂, —CH₂C₆H₅, —CH₂-p-(OH)C₆H₄,—CH₂CH₂CH₂CH₂NH₂, or R₂ and R₃ together form

In accordance with still another embodiment of the present inventionthere is provided a compound having the formula:

and salts thereof.

In accordance with still another embodiment of the present inventionthere is provided a method of forming a nanotube. The method includesdisposing a compound having the formula

in a solution at a sufficient concentration so that the nanotube isformed.

In accordance with still another embodiment of the present inventionthere is provided a nanotube which includes molecules having the formula

wherein X is carbon or nitrogen; n is an integer of, 1, 2, 3, or 4; Y isan amino acid having an amino group covalently bound to an α-carbon ofthe amino acid and the amino group is covalently bound to a carbon ofthe (CH₂)_(n) group; and R₁ is aliphatic; and salts thereof, and anumber of the molecules are arranged relative to one another so as toform a supermacrocycle.

It is therefore an object of the present invention to provide new anduseful compounds for forming nanotubes.

It is another object of the present invention to provide improvedcompounds for forming nanotubes.

It is still another object of the present invention to provide a new anduseful method for forming nanotubes.

It is yet another object of the present invention to provide an improvedmethod for forming nanotubes.

It is still another object of the present invention to provide a new anduseful nanotube.

It is yet another object of the present invention to provide an improvednanotube.

The above and other objects, features, and advantages of the presentinvention will become apparent from the following description andattached drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a representation of a compound of the present invention;

FIG. 2A is a molecular model of a supermacrocycle formed from thecompound shown in FIG. 1;

FIG. 2B is a schematic representation of the supermacrocycle shown inFIG. 2A;

FIG. 3 is a molecular model of a nanotube formed from supermacrocyclesof FIG. 2A;

FIG. 4A is a representation of another compound of the presentinvention;

FIG. 4B is a representation of still another compound of the presentinvention;

FIG. 4C is yet another representation of a compound of the presentinvention;

FIG. 5 is a NMR spectrum of a solution of a compound of the presentinvention;

FIG. 6 is an ESI-MS spectrum of a solution of a compound of the presentinvention;

FIG. 7 is a CD spectrum of solutions of compounds of the presentinvention;

FIG. 8 is an absorbance spectrum of a solution of a compound of thepresent invention;

FIG. 9 is a variable temperature CD spectrum of a solution of a compoundof the present invention;

FIG. 10 is a melting curve of a solution of a compound of the presentinvention;

FIG. 11 is a dynamic light scattering regularization diagram of a asolution of a compound of the present invention; and

FIG. 12 is a large area transmission electron micrograph of nanotubes ofthe present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

While the invention is susceptible to various modifications andalternative forms, a specific embodiment thereof has been shown by wayof example in the drawings and will herein be described in detail. Itshould be understood, however, that there is no intent to limit theinvention to the particular form disclosed, but on the contrary, theintention is to cover all modifications, equivalents, and alternativesfalling within the spirit and scope of the invention as defined by theappended claims.

For a system based on hydrogen bonds to self-assemble in water one hasto balance the enthalpic loss (H-bonds) with a consequent entropic gain(stacking interactions and hydrophobic effect). If preorganized, ionicH-bonds could also add to the enthalpic term. With this in mind, thecompound shown in FIG. 1 (hereinafter referred to as L-module 1) wasdesigned and synthesized with the following features (i) a hydrophobicbase unit possessing the Watson-Crick donor-donor-acceptor (DDA) H-bondarray of guanine and (ii) the acceptor-acceptor-donor (AAD) H-bond arrayof cytosine. Note that arrows pointing away from the molecule denote adonor and the arrows pointing toward the molecule denote an acceptor. Acarbon or a nitrogen atom can be inserted at the position denoted by Xin L-module 1. An appropriate counter ion for L-module 1, or othermodules, can be for example CF₃CO₂ ^(−.) Furthermore, a methyl group(^(A)HNCH₃) was introduced into the structure of L-module 1 so as tominimize the peripheral access of water and enforce the formation of anintramolecular ionic hydrogen bond between the side chain secondaryammonium and the neighboring ring carbonyl (see FIG. 1). However, itshould be understood that other bulky hydrophobic groups can be utilizedin the present invention, including but not limited to other aliphaticor alkyl groups, as long as they perform the aforementioned function,i.e. minimize the peripheral access of water and enforce the formationof an intramolecular ionic hydrogen bond, and do not interfere with theformation of the nanotube. In addition, an ethylene spacer unit linkingthe base component to the chiral center was chosen in order to allow forthe intramolecular ionic H-bond (see FIG. 1). It should be appreciatedthat other spacer units, including but not limited to 1, 3, and 4 carbonatom spacers, can be utilized in the present invention as long as theyallow for the aforementioned intramolecular ionic H-bond and do notinterfere with nanotube formation. An amino acid moiety that dictatesthe supramolecular chirality of the resulting assembly is covalentlybound to the spacer unit as shown in FIG. 1. Any amino acid, includingthe stereoisomers thereof, can be utilized as long as it does notinterfere with nanotube formation. For example, amino acids which may beused in the present invention include, but are not limited to, alanine,arginine, asparagine, aspartate, cysteine, glutamine, glutamate,glycine, histidine, isoleucine, leucine, lysine, methiohine,phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine,hydroxyproline, γ-carboxyglutamate, or o-phosphoserine.

It should be appreciated that modules (i.e. molecules) which possess theabove described characteristics will self-assemble into supermacrocyclesor discrete nanotubular assemblies in water or aqueous solutions.Supermacrocycles are defined herein as being a number of organicmolecules covalently or noncovalently bound together so as to form aring structure. For example, as shown in FIGS. 2A and 2B, onesupermacrocycle these compounds will self-assemble into is a 6-mer. Notethat FIG. 2A is a molecular model of the supermacrocycle L-module 1self-assembles into, while FIG. 2B schematically depicts thesupermacrocycle L-module 1 self-assembles into with each trapezoid 10representing one L-module 1 molecule. With respect to FIG. 2A the thinlines represent the hydrogen bond network and the thick lines highlightunique inter-modular NOE's which are discussed in greater detail below.

It should be appreciated that the process of forming nanotubes with themodules of the present invention is hierarchical. In particular, what ismeant by hierarchical is that the modules of the present invention firstself-assembly into the above discussed supermacrocycles, and then thesupermacrocycles self-assembly into nanotubes. For example, one nanotubeassembly L-module 1 could self-assembly into is shown by way of amolecular model in FIG. 3. In particular, FIG. 3 shows 18supermacrocycles arranged in a tubular fashion with a startinginterplane distance of 4.5 angstroms.

It should also be appreciated that the ability of the modules of thepresent invention to self-assembly into supermacrocycles or discretenanotubular assemblies in water or aqueous solutions is unique inseveral respects. For example, other compounds and methods for formingnanotubes require the formation of supermacrocyclic assemblies inorganic solvents, and in solid and liquid crystalline states. Being ableto form soluble nanotubular assemblies in water or aqueous solutions isan advantage of the present invention since it enhances the ease withwhich the nanotubes can be synthesized. In addition, having solublenanotubular assemblies enhances the ability to utilize the nanotubes inother applications, e.g. photonic-devices.

The following is a general synthetic strategy (i.e. schemes 1-4) whichallows oligomerization and functionalization at virtually any positionof a module (i.e. molecule) having the above discussed designcharacteristics. In particular, these schemes were utilized tosynthesize D and L-module 1 and modules 2, 3, and 4 which are shown inFIGS. 4A, 4B, and 4C, respectively. However, it should be understoodthat this strategy can be extended to the preparation of other modules.In particular, the strategy is amenable to oligomerization usingstandard peptide synthesis chemistry, thereby allowing the formation ofnanotubular assemblies with, for example, predetermined dimensions.

The abbreviations used in each scheme of the synthetic strategy are asfollows: Bn (benzyl); Boc (tert-butyloxycarbonyl); Cbz(benzyloxycarbonyl); DCG (dicyclohexylcarbodiimide); 1,2-DCE(1,2-dichloroethane); dH₂O (deionized NanoPure water); DIEA(diisopropylethylamine); DMAP (4-N,N-dimethylaminopyridine); DME(1,2-dimethoxyethane) DMF (dimethylformamide); EtOH (ethanol); Et₃N(triethylamine); h (hour); NaB(OAc)₃H (sodium triacetoxyborohydride);NMMO (N-methylmorpholine N-oxide); p-TsOH (para-toluenesulfonic acid);rt (room temperature); TFA (trifluoroacetic acid); TFAA (trifluorbaceticanhydride); and THF (tetrahydrofuran).

All the new compounds were characterized by ¹H and ¹³C-NMR, and MS. Highresolution MS and elemental analysis were obtained for key intermediatesand final products. In all the cases the analytical data was inagreement with the structures. Note that each compound is assigned areference number, e.g. 1, 2, 3 and so on, or a letter and a number, e.g.L-17 and so on, which is utilized to identify the compound in thesynthetic strategy. Also note that any compound referenced but notspecifically shown in the general synthetic strategy is discussed belowwith respect to a more detailed discussion of the synthesis of thecompounds. Compounds 2, 17, 19, and 26-28 were prepared according to thereported procedures set forth in Yoneda, Chem. Soc. Perkin Trans. 11976, 16, 1805-1808; Krapcho, Synth. Commun. 1990, 20, 2559-2564;Husbands, J. Med. Chem. 1999, 42, 4446-4455; and Ishikawa, Chem. Pharm.Bull. 1992, 40, 846-850, respectively, all incorporated herein byreference. Barbituric acid, 1,5-diaminopentane, D- andL-N^(α)-Boc-N^(Σ)-Cbz-Lysine, all the reagents and solvents arecommercially available from Aldrich, Novabiochem, Fisher Scientific orAdvanced ChemTech.

Scheme 1. (a) 2-Trimethylsilylethanol, DCC, DMAP, CH₂Cl₂, rt, 22 h, 95%.(b) 10% TFA:CH₂Cl₂, rt, 1.5 h, 95%.

Scheme 2. (a) 1, POCl₃, DMF, reflux, 15 h, 65%. (b) 2, Allylamine,CH₂Cl₂, −78°→−20° C., 7 h, 79%. (c) 3, CH₃NH₂, THF, 0° C.→rt, 7 h, 83%.(d) 4, benzylalcohol, NaH, THF, reflux, 22 h, 75%. (e) 5, (Boc)₂O, Et₃N,DMAP, THF, rt, 18 h, 88%. (f) 6, NH₂OH, MeOH, reflux, 3 h, 92%. (g) 7,TFAA, Et₃N, THF, 0° C.→reflux, 5 h, 79%. (h) 8, Cl(O═C)NCO, CH₂Cl₂, 0°C.→rt, 5 h, 97%. (i) 9, 7N-NH₃, MeOH, rt, 2 h, 59% from 8. (j) 10,(Boc)₂, Et₃N, DMAP, rt, 22 h, 89%. (k) 11, OsO₄, NMMO, Acetone:dH₂O(8:1), rt, 23 h, 98%. (l) 12, NaIO₄, CH₂Cl₂:dH₂O (4:1), rt, 36 h, 86%.(m) L-15, 13, DIEA, 1,2-DCE, NaB(OAc)₃H, rt, 16 h, 72%. (n) L-16,Thioanisole:TFA (6:94), 60 h, 58%. (o) D-15, 13, DIEA, 1,2-DCE,NaB(OAc)₃H, rt, 16 h, 70%. (p) D-16, Thioanisole:TFA (6:94), rt, 86 h,63%, (q) 5-tert-butoxycarbonylamino-1-aminopentane, 13, DIEA, 1,2-DCE,NaB(OAc)₃H, rt, 12 h, 55%. (r) 18, Thioanisole:TFA (6:94), rt, 86 h,77%.

Scheme 3. (a) 19, malononitrile, NaH, DME, rt, 12 h, 88%. (b) 20, Na,EtOH, guanidinium hydrochloride, reflux, 5 h, 74%. (c) 21, (Boc)₂O, THF,DMAP, rt, 12 h, 45%. (d) 22, p-TsOH, MeOH, rt, 5 h, 98%. (e) 23,Dess-Martin reagent, CH₂Cl₂, rt, 2 h, 94%. (f) L-15, 24, DIEA, 1,2-DCE,NaB(OAc)₃H, rt, 12 h, 46%. (g) 25, Thioanisole:TFA (6:94), rt, 12 h,82%.

Scheme 4. (a) 26, urea, Na, CH₃OH, reflux, 5 h, 67%. (b) 27, POCl₃,N,N-dimethylaniline, 70° C.→reflux, 4 h, 71%. (c) 28, benzylalcohol,NaH, THF, reflux, 12 h, 45%. (d) 29, NaIO₄, OsO₄, dioxane:dH₂O (4:1),rt, 12 h, 73%. (e) 30, L-15, DIEA, 1,2-DCE, NaB(OAc)₃H, rt, 12 h, 55%.(f) 31, Thioanisole:TFA (6:94), rt, 96 h, 65%.

As previously mentioned, the following is a more detailed discussion ofthe synthesis of the compounds. The abbreviations used are as follows:AcOH (acetic acid); Bn (benzyl) Boc (tert-butyloxycarbonyl); br(broaden); n-BuOH (1-butanol); Cbz (benzyloxycarbonyl); concd(concentrated); CI-MS (chemical impact mass spectrometry); d (doublet);DCC (dicyclohexylcarbodiimide); 1,2-DCE (1,2-dichloroethane); DCU(dicyclohexylurea); dH₂O (deionized NanoPure water); DIEA(diisopropylethylamine); DMAP (4-N,N-dimethylaminopyridine); DME(1,2-dimethoxyethane) DMF (dimethylformamide); EA (ethylacetate); EI-MS(electron impact mass spectrometry); ESI-MS (electrospray ionizationmass spectrometry); Et₂O (diethylether); EtOH (ethanol); Et₃N(triethylamine); FAB-MS (fast atom bombardment mass spectrometry); h(hour); KIPEG (potassium iodide and polyethyleneglycol matrix forFAB-MS); m (multiplet); min (minute); mp (melting point); NBA(4-nitrobenzylalcohol matrix for FAB-MS); NMMO (N-methylmorpholineN-oxide); NMR (nuclear magnetic resonance); p (pentuplet); PD-MS (plasmadesorption mass spectrometry); p-TsOH (para-toluenesulfonic acid); q(quartet); R_(f) (retention factor); rt (room temperature); s (singlet);sat (saturated); t-Bu (tert-butyl); TFA (trifluoroacetic acid); TFAA(trifluoroacetic anhydride); THF (tetrahydrofuran).

Melting points were recorded on a Thomas Hoover capillary melting pointapparatus (Unimelt). ¹H and ¹³C-NMR spectra were recorded on Varian NMRspectrometers (Gemini 200, Inova 300, Unity 500 or Unity Plus 600 MHz)with the solvent as internal reference. The NMR data is presented asfollows: chemical shift, peak assignment, multiplicity, couplingconstant, integration. The mass spectra were performed at the MassSpectrometry Center of Purdue University. As discussed above compounds2, 17, 19, and 26-28 were prepared according to previously reportedprocedures. Barbituric acid, 1,5-diaminopentane, D andL6-benzyloxycarbonylamino-2-tert-butoxycarbonyl-amino-hexanoic acid, allthe reagents and solvents are commercially available from Aldrich,Novabiochem, Fisher Scientific or Advanced ChemTech. Reagent gradesolvents were distilled under inert atmosphere (N₂) prior to use: CH₂Cl₂was distilled over CaH₂, THF over Na/benzophenone and CH₃OH over Mg. Allthe reactions were performed under N₂ atmosphere.

For column chromatography, commercial solvents were used withoutpurification. Chromatographic supports were silica flash Merck 60(0.040-0.063 mm) or silica gel Merck 60 (0.063-0.2 mm) for gravitychromatography. Silica-coated TLC plates (Merck F 60₂₅₄) were used formonitoring reaction progress and visualizations were made under UV lightor by chemical staining (KMnO₄/dH₂O, phosphomolybdic acid/EtOH, orninhydrin/n-BuOH/AcOH).

Barbituric acid 1 (structure not shown) (10 g, 78.1 mMol) was added to astirred solution of POCl₃ (47 mL, 77.3 g, 504 mMol) and DMF (6 mL, 5.66g, 77.5 mMol) at rt under N₂ atmosphere. The mixture was refluxed for 15h then allowed to cool down to rt. Excess POCl₃ was removed underreduced pressure (high vacuum rotavap), and the resulting viscousmaterial was carefully poured over crushed ice (250 g) while vigorouslystirring. The resulting pale brown precipitate was then filtered anddried under high vacuum. The desired compound 2 (C₅HCl₃N₂O, 10.7 g, 65%)was obtained as yellow crystalline solid upon sublimation (120° C., 0.05mm Hg). R_(f)=0.43 (10% EA/Hex). mp=130° C. ¹H-NMR (300 MHz, CDCl₃) δ(ppm): 10.45 (C₅H, s, 1H).

To a stirred solution of 2 (6.0 g, 28.4 mMol) in CH₂Cl₂ (50 mL),allylamine (3.34 g, 4.26 mL, 56.8 mMol) was slowly added at −78° C.under N₂ atmosphere. The resulting mixture was stirred at −78° C. for 2h then allowed to warm to −20° C. over a period of 7 h. The reaction wasthen quenched with dH₂O (10 mL) and extracted with CH₂Cl₂ (100 mL). Theorganic layer was washed with dH₂O (2×50 mL) and brine (50 mL), thendried over anhydrous Na₂SO₄. After filtration and evaporation of thesolvent (rotavap) the crude product was purified by gravity silica gelchromatography (0 to 2% EA/Hex). The desired compound 3 was obtained asa colorless liquid (C₈H₇Cl₂N₃O, 5.29 g, 79%). R_(f)=0.33 (10% EA/Hex).¹H-NMR (200 MHz, CDCl₃) δ (ppm): 10.25 (C₅H, s, 1H), 9.32 (NHC₆, br, s,1H), 5.94-5.79 (C₇H, m, 1H), 5.25-5.12 (C₈H, m, 2H), 4.20-4.13 (C₆H, m,2H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 190.9 (C₅), 166.46 (C₂), 163.14(C₁), 162.00 (C₄), 132.73 (C₇), 117.99 (C₈), 107.15 (C₃), 44.01 (C₆).Positive ESI-MS: Expected mass for (M+H⁺)/z, 232.00. Observed, 232.1((M+H⁺)/z, 93%).

To a stirred solution of 3 (9.1 g, 39.2 mMol) in THF (100 mL),methylamine (2M-solution in THF, 39.2 mL, 78.4 mMol,) was added at 0° C.under N₂ atmosphere. The reaction mixture was stirred at 0° C. for 1 hthen at rt for 6 h. The reaction was quenched with sat aqueous NH₄Cl (10mL) and the solvent was removed under reduced pressure (rotovap). Thedesired product 4 (C₉H₁₁ClN₄O, 7.36 g, 83%) was obtained as acrystalline white solid after gravity silica gel chromatography.R_(f)=0.22 (SiO₂, 10% EA/Hex). mp=156° C. ¹H-NMR (200 MHz, CDCl₃) δ(ppm): 10.03 (C₅H, s, 1H), 9.37 (NHC₆, br, s, 1H), 6.92 (NHC₉, br, s,1H), 5.95-5.86 (C₇H, m, 1H), 5.28-5.14 (C₈H, m, 2H), 4.18 (C₆H, t, J=5.4Hz, 2H), 3.01 (C₉H, d, J=4.76 Hz, 3H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm):188.83 (C₅), 165.77 (C₃), 162.65 (C₄), 161.94 (C₂), 134.12 (C₈), 116.97(C₇), 102.04 (C₁), 43.44 (C₆), 28.71 (C₉). EI-MS: Expected mass for(M⁺)/z, 226.06 Observed, 226 ((M⁺)/z, 67%), 211 ((M⁺)/z-CH₃, 100%). Highresolution EI-MS: Expected mass for (M⁺)/z, 226.0621 Observed, 226.0624.

Benzyl alcohol (6.68 g, 6.39 mL, 61.8 mMol) was added to a stirredsuspension of 95% NaH (1.79 g, 70.8 mMol) in THF (10 mL) at rt under N₂atmosphere. After 15 min, the solution was cooled to 0° C. then asolution of compound 4 (7.0 g, 30.9 mMol) in THF (40 mL) was added. Themixture was allowed to warm to rt then it was refluxed for 22 h. Themixture was then cooled to 0° C. and carefully quenched with sat NH₄Cl(5 mL). The solvent was removed under reduced pressure (rotovap), andthe residual solid was dissolved in Et₂O, washed with dH₂O (100 mL) andbrine (50 mL) and dried over anhydrous Na₂SO₄. Filtration, evaporationof the solvent under reduced pressure (rotavap) followed by gravitysilica gel chromatography (0 to 5% EA/Hex) yielded 5 as a white solid(C₁₆H₁₈N₄O₂, 7.0 g, 75%). R_(f)=0.51 (30% EA/Hex). mp=56° C. ¹H-NMR (200MHz, CDCl₃) δ (ppm): 10.32 (C₅H, s, 1H), 9.42 (NHC₆, br, s, 1H, majorisomer), 9.25 (NHC₆, br, s, 1H, minor isomer), 7.49-7.37 (C₁₂H—C₁₆H, m,5H), 6.03-5.84 (C₇H, br, m, 1H), 5.46-5.10 (C₈H, C₁₀H, m, 4 H), 4.19(C₆H, br, s, 2H, major isomer), 4.06 (C₆H, br, s, 2H, minor isomer),2.98 (C₉H, d, J=8.5 Hz, 3H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 186.42(C₅), 163.86 (C₃), 163.37 (C₄), 141.82 (C₂), 137.09 (C₁₁), 134.79 (C₈),129.03, 128.93, 127.43 (C₁₂-C₁₆), 116.43 (C₇), 93.92 (C₁), 68.06 (C₁₀),43.32 (C₆), 28.68 (C₉). EI-MS: Expected mass for (M⁺)/z, 298.14.Observed, 298 ((M⁺)/z, 15%), 283 ((M⁺)/z-CH₃, 2.1%), 207 ((M⁺)/z-C₇H₇,27%), 91 ((C₇H₇)⁺, 100%). High resolution EI-MS: Expected mass for(M⁺)/z, 298.1430. Observed, 298.1424.

To a stirred solution of compound 5 (5.0 g, 16.8 mMol), DMAP (1.0 g,8.35 mMol) and THF (50 mL), Et₃N (5.1 g ,7.0 mL, 50.3 mMol) was added atrt under N₂ atmosphere. After stirring for 5 min, Boc₂O (4.39 g, 20.1mMol) was added, and the mixture was stirred at rt for 18 h. Thereaction was quenched with dH₂O (10 mL) followed by removal of thesolvent under reduced pressure (rotovap). The residual solid wasdissolved in EA (300 mL) and washed with 10% aqueous citric acid (50mL), dH₂O (2×50 mL), 5% aqueous NaHCO₃ (50 mL) and brine (50 mL). Afterdrying the organic layer over anhydrous Na₂SO₄, filtration and removalof the solvent under reduced pressure (rotavap), the residue waspurified by gravity silica gel chromatography (0 to 5% EA/Hex). Thedesired compound 6 (C₂₁H₂₆N₄O₄, 5.86 g, 88%) was obtained as a whitecrystalline solid. R_(f)=0.38 (10% EA/Hex). mp=54° C. ¹H-NMR (300 MHz,CDCl₃) δ (ppm): 10.18 (C₅H, s, 1H), 9.26 (NHC₆, br, t, J=5.7 Hz, 1H),7.49-7.36 (C₁₂H-C₁₆H, br, m, 5H), 6.01-5.90 (C₇H, m, 1H), 5.53 (C₁₀H, s,2H), 5.31-5.14 (C₈H, br, m, 2H), 4.23 (C₆H, t, 2H, J=4.8 Hz), 3.43 (C₉H,s, 3H), 1.59 (C₁₉H, s, 9H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 188.09(C₅), 171.89 (C₁₇), 163.16 (C₁), 162.30 (C₂), 154.25 (C₄), 136.79 (C₁₁),134.57 (C₇), 129.03 (C₁₂-C₁₆), 128.68 (C₁₂-C₁₆), 128.54 (C₁₂-C₁₆),116.70 (C₈), 94.75 (C₃), 82.39 (C₁₈), 68.74 (C₁₀), 43.48 (C₆), 35.15(C₉), 28.69 (C₁₉). CI-MS: Expected mass for (M+H⁺)/z, 399.20. Observed,399 ((M+H⁺)/z, 100%), 299 ((M+H⁺)/z-Boc, 7.1%). High resolution EI-MS:Expected for (M⁺)/z, 398.1954. Observed, 398.1943.

To a stirred solution of 6 (4.2 g, 10.6 mMol) in anhydrous methanol (100mL) KHCO₃ (4.23 g, 42.2 mMol) and hydroxylamine hydrochloride (1.47 g,21.1 mMol) were added at rt under N₂ atmosphere. The resulting slurrywas refluxed for 3 h then cooled to rt and quenched with dH₂O (10 mL).The solvent was removed (rotovap) and the residual solid was dissolvedin EA (200 mL), washed with dH₂O (50 mL) and brine (25 mL). The organiclayer was dried over anhydrous Na₂SO₄, filtered, and evaporated todryness (rotavap) to yield compound 7 (C₂₁H₂₇NO₄, 4 g, 92%). Thismaterial was used in the next step without further purification.R_(f)=0.50 (30% EA/Hex). ¹H-NMR (200 MHz, CDCl₃) δ (ppm): 8.58 (C₅H, s,1H), 8.05 (NHC₆, t, J=6.5 Hz, 1H), 7.60 (NOH, br, s, 1H), 7.46-7.34(C₁₂H-C₁₆H, m, 5H), 6.06-5.80 (C₇H, m, 1H), 5.44 (C₁₀H, s, 2H),5.28-5.10 (C₈H, m, 2H), 4.25-4.18 (C₆H, m, 2H), 3.41 (C₉H, s, 3H), 1.89(C₁₉H, s, 9H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 167.63 (C₅), 161.16(C₄), 159.53 (C₂), 155.03 (C₁₇), 146.42 (C₁), 137.25 (C₁₁), 135.30 (C₇),128.95, 128.74 (C₁₂-C₁₆), 116.07 (C₈), 88.54 (C₃), 81.96 (C₁₈), 68.63(C₁₀), 43.85 (C₆), 35.34 (C₉), 28.84 (C₁₉). CI-MS: Expected mass for(M+H⁺)/z, 414.20 Observed, 414 ((M+H⁺)/z, 100%), 396 ((M+H⁺)/z-H₂O,18.3%), 314 ((M+H⁺)/z-Boc, 18.3%). High resolution EI-MS: Expected for(M⁺)/z, 413.2063. Observed, 413.2050.

Compound 7 (4.0 g, 9.66 mMol), Et₃N (4.03 mL, 2.93 g, 29.0 mMol) and THF(50 mL) were cooled to 0° C. then TFAA (2.0 mL, 3.04 g, 14.5 mMol) wasslowly added. After stirring for 15 min, the mixture was allowed to warmto rt then it was refluxed for 5 h. After cooling down to rt thereaction was quenched with dH₂O (10 mL) and the solvent was removedunder reduced pressure (rotovap). The residual solid was dissolved in EA(300 mL), washed with dH₂O (2×50 mL), 10% aqueous citric acid (25 mL),dH₂O (50 mL), 5% aqueous NaHCO₃ (50 mL) and brine (50 mL). The organiclayer was dried over anhydrous Na₂SO₄, filtered, and evaporated todryness under reduced pressure (rotavap). Compound 8 was obtained as awhite solid (C₂₁H₂₅N₅O₃, 3.3 g, 79% from 6) after gravity silica gelchromatography (3% EA/Hex). R_(f)=0.64 (30% EA/Hex). mp=68° C. ¹H-NMR(300 MHz, CDCl₃) δ (ppm): 7.49-7.35 (C₁₂H-C₁₆H, m, 5H), 5.91-5.83 (C₇H,m, 1H), 5.78 (NHC₆, t, J=5.6 Hz, 1H), 5.51 (C₁₀H, s, 2H), 5.27 (C₈H, d,J=17.1 Hz, 1H), 5.19 (C₈H, d, J=10.1 Hz, 1H), 4.18 (C₆H, t, J=5.7 Hz,2H), 3.40 (C₉H, s, 3H), 1.58 (C₁₉H, s, 9H). ¹³C-NMR (75 MHz, CDCl₃) δ(ppm): 170.62 (C₁₇), 164.31 (C₁), 161.05 (C₂), 153.80 (C₄), 136.26(C₁₁), 134.14 (C₇), 128.76 (C₁₂-C₁₆), 128.44 (C₁₂-C₁₆), 128.25(C₁₂-C₁₆), 117.20 (C₈), 115.05 (C₅), 82.27 (C₁₈), 69.06 (C₁₀), 68.83(C₃), 43.90 (C₆), 34.94(C₉), 28.47 (C₁₉). CI-MS: Expected mass for(M+H⁺)/z, 396.20. Observed, 396 ((M+H⁺)/z, 100%), 296 ((M+H⁺)/z-Boc,32%). High resolution EI-MS: Expected for (M⁺)/z, 395.1957. Observed,395.1961.

To a solution of compound 8 (15.2 g, 38.4 mMol) in CH₂Cl₂ (300 mL),N-chlorocarbonylisocyanate (Gorbatenko, Tetrahedron 1993, 49, 3227-3257incorporated herein by reference) (8.09 g, 6.2 mL, 76.7 mMol) was addeddropwise at 0° C. over a period of 15 min under N₂ atmosphere. Afterstirring for 2 h at 0° C., the mixture was allowed to warm to rt and wasstirred for an additional 3 h. The reaction mixture was cooled to 0° C.and carefully quenched with dH₂O (10 mL, exothermic reaction!) followedby 5% aqueous NaHCO₃ (10 mL). The product was extracted with chloroform(1500 mL) and the resulting organic layer was washed with dH₂O (2×100mL), brine (100 mL) and dried over anhydrous Na₂SO₄. Filtration andevaporation of the organic solvents under reduced pressure (rotavap)yielded 9 (C₂₂H₂₆N₆O₄, 16.4 g, 97%) as a viscous liquid, which was usedin the next step without further purification. A small sample of thiscompound was purified by gravity silica gel chromatography (30% EA/Hex)to yield 9 as an analytically pure white solid. R_(f)=0.16 (30% EA/Hex).mp=98° C. ¹H-NMR (200 MHz, CDCl₃) δ (ppm): 7.46 (C₁₂H—C₁₆H, br, m, 5H),5.94-5.75 (C₇H, m, 1H), 5.51 (C₁₀H, s, 2H), 5.18 (C₈H, t, J=21.2 Hz,2H), 4.92 (C₆H, d, J=5.4 Hz, 2H), 3.41 (C₉H, s, 3H), 1.54 (C₁₉H, s, 9H).¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 172.54 (C₁₇), 163.51 (C₁), 159.08 (C₂),155.86 (C₄), 152.95 (C₂₀), 135.70 (C₁₁), 133.82 (C₇), 129.14-128.93,128.21 (C₁₂-C₁₆), 117.34 (C₈), 114.44 (C₅), 83.90 (C₁₈), 70.18 (C₁₀),48.51 (C₆), 34.99 (C₉), 28.51 (C₁₉). CI-MS: Expected mass for (M+H⁺)/z,439.20. Observed, 439 ((M+H⁺)/z, 2.8%), 396 ((M+H⁺)/z-CONH₂, 160%), 296((M+H⁺)/z-(Boc+CONH₂), 39%). High resolution EI-MS: Expected mass for(M⁺)/z, 438.2016. Observed, 438.2027.

Compound 9 (16.4 g, 37.3 mMol) was stirred in 7 N NH₃ in CH₃OH (450 mL)under N₂ atmosphere at rt for 2 h. Excess CH₃OH was removed underreduced pressure (rotovap) and the desired compound 10 was obtained as awhite solid (C₂₂H₂₆N₆O₄, 9.94 g, 59% from 8) after gravity silica gelchromatography (50 to 100% Hex/EA, then 0 to 3% CH₃OH/CHCl₃). R_(f)=0.26(EA). mp=192° C. ¹H-NMR (300 MHz, CDCl₃) δ (ppm): 8.00 (NHC₅, br, s,1H), 7.51-7.41 (C₁₂H-C₁₆H, m, 5H), 7.15 (NHC₅, s, 1H), 6.02-5.91 (C₇H,br, m, 1H), 5.66 (C₁₀H, s, 2H), 5.27 (C₈H, d, J=17.2 Hz, 1H), 5.18 (C₈H,d, J=10.2 Hz, 1H), 4.87 (C₆H, d, J=5.6 Hz, 2H), 3.47 (C₉H, s, 3H), 1.61(C₁₉H, s, 9H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 167.04 (C₁₇), 161.72(C₁), 161.09 (C₂), 160.83 (C₅), 156.37 (C₄), 153.53 (C₂₀), 135.58 (C₁₁),132.99 (C₇), 129.27, 129.07 (C₁₂-C₁₆), 117.80 (C₈), 86.65 (C₃), 82.97(C₁₈), 70.31 (C₁₀), 45.01 (C₆), 35.26 (C₉), 28.64 (C₁₉). CI-MS: Expectedmass for (M+H⁺)/z, 439.20. Observed, 439 ((M+H⁺)/z, 100%), 339((M+H⁺)/z-Boc, 29%). Positive high resolution CI-MS: Expected mass for(M+H⁺)/z, 439.2094. Observed, 439.2096.

To a solution of compound 10 (2.5 g, 5.69 mMol), DMAP (0.7 g, 5.69 mmol)and Et₃N (4.8 mL, 3.5 g, 34.2 mMol) in THF (50 mL), Boc₂O (3.72 g, 17.1mMol) was added under N₂ atmosphere. After stirring at rt for 22 h, thereaction was quenched with dH₂O (10 mL) and the solvent was removedunder reduced pressure (rotovap). The residual solid was dissolved in EA(300 mL), washed with dH₂O (100 mL), 10% aqueous citric acid (50 mL),dH₂O (2×50 mL), 5% aqueous NaHCO₃ (50 mL) and brine (50 mL), then driedover anhydrous Na₂SO₄. After filtration and removal of the solvent underreduced pressure (rotavap) the residual solid was purified by gravitysilica gel chromatography (5 to 20% EA/Hex) to yield compound 11(C₃₂H₄₂N₆O₈, 3.25 g, 89%) as a white foam. R_(f)=0.66 (50% EA/Hex).mp=78° C. ¹H-NMR (200 MHz, CDCl₃) δ (ppm): 7.48-7.44 (C₁₂H-C₁₆H, m, 5H),6.07-5.87 (C₇H, m, 1H), 5.58 (C₁₀H, s, 2H), 5.24 (C₈H, t, J=12.4 Hz,2H), 4.94 (C₆H, d, J=9.2 Hz, 2H), 3.46 (C₉H s, 3H), 1.59 (C₁₉H, s, 9H),1.35 (C₂₃H and C₂₆H, s, 18H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 165.96(C₁₇), 161.37 (C₂₄ or C₂₂), 161.06 (C₂₂ or C₂₄), 160.67 (C₁), 155.60(C₄), 152.88 (C₂₀), 149.41 (C₂, C₅), 135.19 (C₁₁), 131.48 (C₇), 128.85,128.72 (C₁₂-C₁₆), 118.26 (C₈), 93.08 (C₃), 83.83 (C₂₃, C₂₆), 83.23(C₁₈), 70.35 (C₁₀), 45.49 (C₆) 35.16 (C₉), 28.35 (C₁₉), 28.05 (C₂₃,C₂₆). EI-MS: Expected mass for (M+H⁺)/z, 638.31. Observed 639.8((M+H⁺)/z, 89%). High resolution EI-MS: Expected for (M⁺)/z, 638.3064.Observed, 638.3057.

To a stirred solution of compound 11 (3.2 g, 5.01 mMol) in acetone:dH₂O(8:1, 90 mL) was added 50% aqueous NMMO (1.17 g, 2.4 mL, 10.0 mMol) atrt. After stirring for 5 min, OsO₄ (0.1 M solution in t-BuOH, 2.5 mL,0.25 mMol) was added dropwise over a period of 5 min. The resultingbrown solution was stirred at rt for 23 h then quenched with aqueoussodium sulfite until all the excess OsO₄ was destroyed (brown solutionturns colorless). Diol 12 was extracted in CHCl₃ (250 mL) and washedwith dH₂O (2×10 mL) and brine (25 mL). The organic layer was dried overanhydrous Na₂SO₄, filtered, and evaporated to dryness under reducedpressure (rotavap). Crystallization from EA yielded compound 12 as awhite solid (C₃₂H₄₄N₆O₁₀, 3.31 g, 98%). R_(f)=0.15 (50% EA/Hex). mp=136°C. ¹H-NMR (300 MHz, CDCl₃) δ (ppm): 7.46 (C₁₂H-C₁₆H, br, m, 5H), 5.61(C₁₀H, s, 2H), 4.63 (C₆H, d, J=4.5 Hz, 2H), 4.18 (C₇H, m, 1H), 4.09(C₇OH, d, J=5.6 Hz, 1H), 3.60 (C₈H, m, 2H), 3.51 (C₉H, s, 3H), 3.28(C₈OH, t, J=7.2 Hz), 1.62 (C₁₉H, s, 9H,), 1.39 (C₂₃H, C₂₆H, s, 18H).¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 166.33 (C₁₇), 162.24 (C₁), 161.25.(C₂₁), 160.00 (C₂₄), 157.36 (C₄), 152.81 (C₂₀), 149.71 (C₂), 149.63(C₅), 135.15 (C₁₁), 129.14, 128.51 (C₁₂-C₁₆), 93.76 (C₃), 84.46 (C₂₂,C₂₅), 84.34 (C₁₈), 71.18 (C₇), 7.0.77 (C₈), 63.79 (C₁₀), 45.90 (C₆),35.21 (C₉), 28.52 (C₁₉), 28.29 (C₂₆, C₂₃). CI-MS: Expected mass for(M+H⁺)/z, 673.31. Observed 673.8 ((M+H⁺)/z, 100%), 573.8 ((M+H⁺)/z-Boc,23%). EI-MS: Expected mass for (M⁺)/z, 672.31. Observed 711 ((M+K⁺)/z,11%), 672.2 ((M⁺)/z, 33%), 373 ((M+H⁺)/z-3Boc, 93%). High resolutionEI-MS: Expected mass for (M⁺)/z, 672.3119. Observed, 672.3131.

A solution of compound 12 (12.0 g, 17.8 mMol) in CH₂Cl₂/dH₂O (4:1, 250mL) and sodium periodate (7.63 g, 35.7 mMol) was stirred at rt for 36 h.The mixture was then filtered through a pad of celite and washed withCH₂Cl₂ (2×200 mL). Separation and evaporation of the organic layer underreduced pressure (rotavap) followed by gravity silica gel chromatography(5 to 30% EA/Hex) yielded compound 13 (C₃₁H₄₀N₆O₉, 9.82 g, 86%) as awhite foam. R_(f)=0.48 (30% EA/Hex). mp=152° C. ¹H-NMR (300 MHz, CDCl₃)δ (ppm): 9.58 (C₇H, d, J=1.2 Hz, 1H), 7.39-7.29 (C₁₂H-C₁₆H, br, m, 5H),5.53 (C₁₀H, s, 2H), 5.09 (C₆H, s, 2H), 3.34 (C₉H, s, 3H), 1.49 (C₁₉H, s,9H), 1.27 (C₂₃H, C₂₆H, s, 18H). ¹³C-NMR (75 MHz, CDCl₃) δ (ppm): 193.96(C₇), 165.82 (C₁₇), 161.24 (C₂₁), 161.16 (C₂₄), 160.88 (C₁), 155.57(C₄), 152.51 (C₂₀), 149.18 (C₂, C₅), 134.93 (C₁₁), 128.83, 128.79,128.62 (C₁₂-C₁₆), 92.99 (C₃), 84.04 (C₂₂, C₂₅), 83.39 (C₁₈), 70.42(C₁₀), 52.20 (C₆), 35.05 (C₉), 28.23 (C₁₉), 27.94 (C₂₆, C₂₃). PD-MS:Expected mass for (M+H⁺)/z, 641.29. Observed, 678.7 ((M+H⁺)/z+K⁺), 641.7((M+H⁺)/z), 613.0 ((M+H⁺)/z-CO). High resolution CI-MS: Expected for(M+H⁺)/z, 641.2935. Observed, 641.2945.

To a stirred solution of6-benzyloxycarbonylamino-2-tert-butoxycarbonyl-L-amino-hexanoic acid(5.0 g, 13.4 mMol), DMAP (0.161 g, 1.32 mMol) and2-trimethylsilylethanol (1.55 g, 13.1 mMol) in CH₂Cl₂ (50 mL), DCC (2.98g, 14.5 mMol) was added at 0° C. under N₂ atmosphere. After 15 min at 0°C., the mixture was stirred at rt for an additional 22 h. The solutionwas filtered to remove the DCU formed and concentrated under reducedpressure (rotavap). L-14 was obtained as a colorless viscous liquid(C₂₄H₄₀N₂O₆Si, 6.0 g, 95%) after gravity silica gel chromatrography (0to 5% EA/Hex). R_(f)=0.57 (30% EA/Hex). ¹H-NMR (200 MHz, CDCl₃) δ (ppm):7.39 (C₃₉H-C₄₃H, s, 5H), 5.14 (C₃₇H, s, 2H), 4.88 (C₃₁H, br, s, 1H),4.23 (C₂₉H, t, J=9.0 Hz, 2H), 3.18 (C₃₅H, q, J=9.0 Hz, 1H), 1.91-1.11(C₃₂H, C₃₃H, C₃₄H, br, m, 6H), 1.46 (C₄₆H s, 9H), 1.00 (C₂₈H, t, J=9.0Hz, 2H), 0.06 (C₂₇H, s, 9H). ¹³C-NMR (50 MHz, CDCl₃) δ (ppm): 172.70(C₃₀), 156.33 (C₄₄), 155.31 (C₃₆), 136.46 (C₃₈), 128.26, 127.87, 127.82(C₃₉-C₄₃), 79.50 (C₄₅), 66.31 (C₃₇), 63.43 (C₂₉), 53.12 (C₃₁), 40.41(C₃₅), 32.11 (C₃₂), 29.15 (C₃₃), 28.12 (C₄₆), 22.22 (C₃₄), 17.15 (C₂₈),−1.71 (C₂₇). Positive ESI-MS: Expected mass for (M+H⁺)/z, 481.27.Observed, 480.6 ((M+H⁺)/z, 91%), 503.1 ((M+Na⁺)/z, 100%). Highresolution EI-MS: Expected for (M+H⁺)/z, 480.2656. Observed, 480.2662.

To a stirred solution of6-benzyloxycarbonylamino-2-tert-butoxycarbonyl-D-amino-hexanoic acid(17.5 g, 46.0 mMol), DMAP (0.56 g, 4.60 mMol) and2-trimethylsilylethanol (5.44 g, 46.0 mMol) in CH₂Cl₂ (250 mL), DCC(10.44 g, 50.6 mMol) was added at 0° C. under N₂ atmosphere. After 15min at 0° C., the solution was stirred at rt for an additional 7.5 h.The solution was filtered to remove the DCU formed and concentratedunder reduced pressure (rotavap). D-14 was obtained as a colorlessviscous liquid (C₂₄H₄₀N₂O₆Si, 20.23 g, 92%) after gravity silica gelchromatrography (0 to 5% EA/Hex). R_(f)=0.57 (30% EA/Hex). ¹H-NMR (300MHz, CDCl₃) δ (ppm): 7.16 (C₃₉H-C₄₃H, br, s, 5H), 5.57 (C₃₇H, s, 2H),5.39 (C₃₁NH, d, J=8.0 Hz, 1H), 4.92 (C₃₁H, s, 1H), 4.05 (C₂₉H, t, J=9.0Hz, 2H), 2.99 (C₃₅H, d, J=5.7 Hz, 2H), 1.61-1.13 (C₃₂H, C₃₃H, C₃₄H, m,6H,), 1.28 (C₄₆H, s, 9H), 0.84 (C₂₈H, t, J=9.0 Hz, 2H), −0.102 (C₂₇H, s,9H). ¹³C-NMR (75 MHz, CDCl₃) δ (ppm): 172.94 (C₃₀), 155.66 (C₄₄), 155.60(C₃₆), 136.78 (C₃₈), 128.35, 127.93, 127.87 (C₃₉-C₄₃), 79.38 (C₄₅),66.27 (C₃₇), 63.40 (C₂₉), 53.41 (C₃₁), 40.44 (C₃₅), 31.95 (C₃₂), 29.28(C₃₃), 28.27 (C₄₆), 22.43 (C₃₄), 17.25 (C₂₈), −1.56 (C₂₇). CI-MS:Expected mass for (M+H⁺)/z, 481.27. Observed, 481 ((M+H⁺)/z, 10%), 381((M+H⁺)/z-Boc, 100%).

A solution of compound L-14 (1.4 g, 2.91 mMol) in 10% TFA/CH₂Cl₂ (50 mL)was stirred at rt under N₂ atmosphere for 1.5 h then the excess TFA andsolvent was removed under reduced pressure (rotavap). The resultingviscous material was dissolved in CH₂Cl₂ (50 mL) and concentrated onceagain under reduced pressure (rotavap) to yield the TFA salt of compoundL-15 (C₁₉H₃₂N₂O₄Si—CF₃CO₂H, 1.35 g, 95%) as a colorless foam. Thiscompound was used in the next step without further purification.R_(f)=0.5 (10% CH₃OH/CHCl₃). ¹H-NMR (200 MHz, CDCl₃ with a drop ofCH₃OD) δ (ppm): 7.30 (C₃₉H-C₄₃H, m, 5H), 5.03 (C₃₇H, s, 2H), 4.26 (C₂₉H,t, J=9.0 Hz, 2H), 3.92 (C₃₁H, br, t, J=3.0 Hz, 1H), 3.12 (C₃₅H, br, s,2H), 1.89 (C₃₂H, br, s, 2H), 1.46 (C₃₃H, C₃₄H, br, s, 4H), 1.01 (C₂₈H,t, J=9.2 Hz, 2H), 0.01 (C₂₇H, s, 9H). ¹H-NMR (300 MHz, CDCl₃ with a dropof TFA) δ (ppm): 8.60-8.20 (TFA, br, s, 6H), 7.31 (C₃₉H-C₄₃H, s, 5H),5.06 (C₃₇H, s, 2H), 4.32-4.26 (C₂₉H, m, 2H), 3.98 (C₃₁H, br, s, 1H),3.15 (C₃₅H, br, s, 2H), 1.95 (C₃₂H, br, s, 2H), 1.80-1.20 (C₃₃H, C₃₄H,m, 4H), 1.05-1.00 (C₂₈H, m, 2H), 0.04 (C₂₇H, s, 9H). ¹³C-NMR (75 MHz,CDCl₃) δ (ppm): 169.79 (C₃₀), 157.87 (C₃₆), 136.45 (C₃₈), 128.69,128.33, 127.90 (C₃₉-C₄₃), 67.19 (C₃₇), 65.87 (C₂₉), 53.44 (C₃₁), 40.36(C₃₅), 29.88 (C₃₂), 29.13 (C₃₄), 21.81 (C₃₃), 17.37 (C₂₈), −1.88 (C₂₇).CI-MS: Expected mass for (M+H⁺)/z, 381.21. Observed, 381.10 ((M+H⁺)/z,26%). High resolution ESI-MS: Expected mass for (M+H⁺)/z, 381.2210.Observed, 381.2214.

A solution of compound D-14 (1.6 g, 3.30 mMol) in 10% TFA/CH₂Cl₂ (50 mL)was stirred at rt under N₂ atmosphere for 1.5 h then the excess TFA andsolvent was removed under reduced pressure (rotavap). The resultingviscous material was dissolved in CH₂Cl₂ (50 mL) and concentrated onceagain under reduced pressure (rotavap) to yield the TFA salt of compoundD-15 (C₁₉H₃₂N₂O₄Si—CF₃CO₂H, 1.59 g, 96%) as a colorless foam. Thiscompound was used in the next step without further purification.R_(f)=0.5 (10% CH₃OH/CHCl₃). ¹H-NMR (300 MHz, CDCl₃) δ (ppm): 8.85 (TFA,br, s, 4H), 8.20 (NHC₃₁, br, s, 2H), 7.35-7.30 (C₃₉H—C₄₃H, m, 5H), 5.09(C₃₇H, s, 2H), 4.32 (C₂₉H, t, J=4.0, 2H), 4.02 (C₃₁H, br, m, 1H), 3.18(C₃₅H, br, s, 2H), 1.98 (C₃₂H, br, s, 2H), 1.57-1.46 (C₃₃H, C₃₄H, m,4H), 1.07 (C₂₈H, t, J=4.4 Hz, 2H), 0.07 (C₂₇H, s, 9H). ¹³C-NMR (75 MHz,CDCl₃) δ (ppm): 169.79 (C₃₀), 157.87 (C₃₆), 136.45 (C₃₈), 128.74,128.42, 127.95 (C₃₉-C₄₃), 67.19 (C₃₇), 65.87 (C₂₉), 53.44 (C₃₁), 40.36(C₃₅), 29.88 (C₃₂), 29.13 (C₃₄), 21.87 (C₃₃), 17.37 (C₂₈), −1.88 (C₂₇).CI-MS: Expected mass for (M+H⁺)/z, 381.21. Observed, 381 ((M+H⁺)/z,26%).

To a stirred solution of L-15 (TFA salt, 0.8 g, 1.53 mmol) and DIEA(0.53 mL, 0.39 g, 3.07 mmol) in 1,2-DCE (30 mL), a solution of compound13 (0.98 g, 1.53 mmol) in 1,2-DCE (20 mL) was added at rt under N₂atmosphere. After stirring for 15 min at rt, solid NaB(OAC)₃H (0.48 g,2.3 mmol) was added (Abdel-Magid, J. Org. Chem. 1996, 61, 3849-3862incorporated herein by reference). The resulting solution was stirredfor 16 h at rt then quenched with dH₂O (10 mL). The aqueous layer wasextracted with CHCl₃ (200 mL) and the organic layers were combined,washed with 10% aqueous citric acid (50 mL), dH₂O (2×50 mL) and brine(50 mL), then dried over anhydrous Na₂SO₄. Filtration and evaaporationof the solvent under reduced pressure (rotavap) followed by gravitysilica gel chromatography (5 to 20% EA/Hex) yielded compound L-16 as acolorless foam (C₅₀H₇₂N₈O₁₂Si, 1.11 g, 72%). R_(f)=0.25 (30% EA/Hex).¹H-NMR (500 MHz, CDCl₃) δ (ppm): 7.38-7.27 (C₁₅H and C₁₃H, m, 2H),7.26-7.19 (C₃₉H-C₄₃H, C₁₂H, C₁₆H, C₁₄H, br, m, 8H), 5.49 (C₁₀H, s, 2H),5.09 (NHC₃₅, m, 1H,), 4.99 (C₃₇H, s, 2H), 4.35 (C₆H, t, J=6.5 Hz, 2H),4.10 (C₂₉H, t, J=8.5 Hz, 2H), 3.40 (C₉H, s, 3H), 3.15 (C₇H, t, J=6.5 Hz,1H), 3.05-3.01 (C₇H, C₃₅H, br, m, 3H), 2.75 (C₃₁H, br, q, 1H), 1.52(C₁₉H, s, 9H), 1.39-1.16 (C₃₂H, C₃₃H, C₃₄H, br, m, 6H), 1.25 (C₂₃H,C₂₆H, s, 18H), 0.91 (C₂₈H, t, J=8.5 Hz, 2H), −0.03 (C₂₇H, s, 9H).¹³C-NMR (125 MHz, CDCl₃) δ (ppm): 175.64 (C₃₀), 165.87 (C₁₇), 161.37(C₂₁), 161.14 (C₂₄), 160.44 (C₁), 156.60 (C₂), 156.04 (C₄), 152.78(C₂₀), 149.53 (C₅), 136.95 (C₃₈), 135.10 (C₁₁), 128.86, 128.74, 128.73,128.61, 128.55 (C₁₂-C₁₆), 128.30, 128.20, 128.13 (C₃₉-C₄₃), 93.04 (C₃),83.89 (C₂₂, C₂₅), 83.19 (C₁₈), 70.22 (C₁₀), 66.55 (C₃₇), 63.04 (C₂₉),61.21 (C₃₁), 45.40 (C₆), 43.07 (C₇), 41.00 (C₃₅), 35.05 (C₉), 33.12(C₃₂), 29.79 (C₃₄), 28.29 (C₁₉), 27.97 (C₂₃, C₂₆), 23.15 (C₃₃), 17.66(C₂₈), −1.30 (C₂₇). PD-MS: Expected mass for (M+H⁺)/z, 1005.5. Observed,1007 ((M+H⁺)/z), 805 ((M+H⁺)/z-2Boc), 705 ((M+H⁺)/z-3Boc). PositiveESI-MS. Expected mass for (M+H⁺)/z, 1005.5. Observed, 1005:3 ((M+H⁺)/z,100%), 805.3 ((M+H⁺)/z-2Boc, 9%), 705.4 ((M+H⁺)/z-3Boc, 7%). Positivehigh resolution ESI-MS: Expected mass for (M+H⁺)/z, 1005.5117. Observed,1005.5101.

To a stirred solution of D-15 (TFA salt, 0.70 g, 1.34 mMol) and DIEA(0.60 mL, 0.45 g, 3.44 mMol) in 1,2-DCE (25 mL), a solution of compound13 (0.86 g, 1.34 mMol) in 1,2-DCE (25 mL) was added at rt under N₂atmosphere. After stirring for 15 min at rt, solid NaB(OAc)₃H (0.54 g,2.58 mMol) was added. The resulting solution was stirred for 16 h at rtthen quenched with dH₂O (10 mL). The aqueous layer was extracted withCHCl₃ (200 mL) and the organic layers were combined, washed with 10%aqueous citric acid (50 mL), dH₂b (2×50 mL) and brine (50 mL), thendried over anhydrous Na₂SO₄. Filtration and evaporation of the solventunder reduced pressure (rotavap) followed by gravity silica gelchromatography (5 to 20% EA/Hex) yielded compound D-16 as a colorlessfoam (C₅₀H₇₂N₈O₁₂Si, 0.95 g, 70%). R_(f)=0.25 (30% EA/Hex). mp=52° C.¹H-NMR (500 MHz, CDCl₃) δ (ppm): 7.39-7.27 (C₁₅H, C₁₃H, m, 2H),7.26-7.18 (C₃₉H-C₄₃H, C₁₂H, C₁₆H, C₁₄H, br, m, 8H), 5.50 (C₁₀H, s, 2H),5.09 (C₃₅NH, m, 1H), 5.00 (C₃₇H, s, 2H), 4.36 (C₆H, t, J=6.5 Hz, 2H),4.10 (C₂₉H, t, J=9 Hz, 2H), 3.40 (C₉H, s, 3H), 3.16 (C₇H, t, J=6.5 Hz,1H), 3.06-2.97 (C₇H, C₃₅H, br, m, 3H), 2.76 (C₃₁H, br, q, 1H), 1.52(C₁₉H, s, 9H), 1.39-1.13 (C₃₂H, C₃₃H, C₃₄H, br, m, 6H), 1.26 (C₂₃H,C₂₆H, s, 18H), 0.92 (C₂₈H, t, J=8.5 Hz, 2H), −0.03 (C₂₇H, s, 9H).¹³C-NMR (125 MHz, CDCl₃) δ (ppm): 175.64 (C₃₀), 165.87 (C₁₇), 161.37(C₂₁), 161.14 (C₂₄), 160.45 (C₁), 156.60 (C₂), 156.04 (C₄), 152.77(C₂₀), 149.54 (C₅), 136.96 (C₃₈), 135.11 (C₁₁), 128.86, 128.75, 128.73,128.61, 128.55 (C₁₂-C₁₆), 128.30, 128.20, 128.13 (C₃₉-C₄₃), 93.04 (C₃),83.89 (C₂₂), 83.87 (C₂₅), 83.19 (C₁₈), 70.23 (C₁₀), 60.55 (C₃₇), 63.04(C₂₉), 61.22 (C₃₁), 45.41 (C₆), 43.07 (C₇), 41.01 (C₃₅), 35.05 (C₉),33.12 (C₃₂), 29.80 (C₃₄), 28.29 (C₁₉), 27.98 (C₂₃, C₂₆), 23.15 (C₃₃),17.67 (C₂₈), −1.30 (C₂₇). Positive ESI-MS: Expected mass for (M+H⁺)/z,1005.5. Observed, 1005.2 ((M+H⁺)/z, 100%), %), 805.2 ((M+H⁺)/z-2Boc,30%), 705.3 ((M+H⁺)/z-3Boc, 10%).

To a stirred solution of 1,5-diaminopentane (4.5. g, 44.0 mMol) indioxane (15 mL), a solution of Boc₂O (1.23 g, 5.5 mMol) in dioxane (15mL) was slowly added at rt over a period of 2.5 h. After stirring for 22h the solvent was removed under reduced pressure (rotovap), dH₂O (25 mL)was added and the resulting precipitate (undesiredbis-N-Boc-2,5-diaminopentane) was filtered. The filtrate was extractedwith CH₂Cl₂ (3×50 mL) and the combined organic layers were evaporatedunder reduced pressure. Compound 17 (C₁₀H₂₂N₂O₂, 0.88 g, 80%) wasobtained as a viscous liquid, which was used in the next step withoutfurther purification. R_(f)=0.47 (10% CH₃OH/CHCl₃). ¹H-NMR (200 MHz,CDCl₃) δ (ppm): 4.90 (NHC₃₅, br, s, 1H), 3.03 (C₃₅H, br, s, 2H), 2.62(C₃₁H, br, s, 2H), 1.37 (C₃₈H, C₃₂H, C₃₃H, C₃₄H, br, s, 15H,), 1.08(NHC₃₁, br, s, 2H).

To a stirred solution of 17 (0.20 g, 1.0 mMol) and compound 13 (0.64 g,1.0 mMol) in 1,2-DCE (30 mL), Na(AcO)₃BH (0.32 g, 1.5 mMol) was added atrt under N₂ atmosphere. The resulting solution was stirred 12 h, at rtthen quenched with dH₂O (10 mL). The aqueous layer was extracted withCHCl₃ (45 mL) and the organic layers were combined and washedsuccessively with 10% aqueous citric acid (50 mL), dH₂O (2×50 mL) andbrine (50 mL), then dried over anhydrous Na₂SO₄. Filtration andevaporation of the solvent under reduced pressure (rotavap) followed bygravity silica gel chromatography (5 to 20% EA/Hex) yielded compound 18as a colorless foam (C₄₁H₆₂N₈O₁₀, 0.46 g, 55%). R_(f)=0.44 (2%CH₃OH/CHCl₃). ¹H-NMR (300 MHz, CDCl₃) δ (ppm): 7.48-7.36 (C₁₂H-C₁₆H, m,5H), 5.59 (C₁₀H, s, 2H), 4.59 (NHC₃₁, br, s, 1H), 4.48 (C₆H, t, 2H,J=6.6 Hz), 3.49 (C₉H, s, 3H), 3.01 (C₇H, t, J=6.6 Hz, 2H), 2.65 (C₃₁H,t, J=6.9 Hz, 2H), 1.61 (C₃₈H, s, 9H), 1.46 (C₁₉H, s, 9H), 1.35 (C₂₃H andC₂₆H, s, 18H), 0.88 (C₃₄H, q, J=14.1 and 6.0 Hz, 2H). Positive ESI-MS:Expected mass for (M+H⁺)/z, 827.47. Observed, 827.3 ((M+H⁺)/z, 100%),727.2 ((M+H⁺)/z,-Boc, 8%), 627.3 ((M+H⁺)/z,-2Boc, 13%), 527.3((M+H⁺)/z,-3Boc, 17%), 427.3 ((M+H⁺)/z,-4Boc, 31%). Negative ESI-MS:Expected mass for (M−H⁺)/z, 825.46. Observed, 825.4 ((M−H⁺)/z, 73%).Positive high resolution ESI-MS: Expected mass for (M+H⁺)/z, 827.4667.Observed, 827.4659.

A 94% solution of TFA/thioanisole (10 mL) was added to L-16 (0.48 g,0.48 mMol) at rt under N₂ atmosphere. After stirring for 60 h at rt,Et₂O (50 mL) was added. The precipitate formed was centrifuged andwashed with Et₂O (3×10 mL), redissolved in TFA (15 mL), reprecipitatedwith Et₂O (20 mL) then washed with Et₂O (2×10 mL) and CH₃OH (10 mL).L-Module 1 was obtained as a white solid(C₁₅H₂₄N₈O₄—(CF₃CO₂H)_(2.5)—(H₂O)_(1.5), 0.168 g, 45%). mp>298° C.(decomposed). ¹H-NMR (600 MHz, 90% H₂O/D₂O) δ (ppm): 9.15 (C₅NH₂, br, s,2H), 8.34 (C₄NH, br, s, 1H), 7.57 (C₉NH, br, m, 1H), 7.39 (C₃₅NH, br, s,3H, protonated ammonium), 3.81 (C₃₁H, t, J=5.0 Hz, 1H), 3.36 (C₇H, t,J=6.0 Hz, 2H), 2.90 (C₉H, br, m, 3H), 2.85 (C₃₅H, br, m, 2H), 1.84-1.79(C₃₂H, m, 2H), 1.55 (C₃₄H, br, s, 2H), 1.36 (C₃₃H, br, m, 1H), 1.29(C₃₃H, br, m, 1H). C₆H was suppressed with the solvent peak. ¹H-NMR(DMSO-d₆, 500 MHz) δ (ppm): 12.60 (C₃₀OH or C₃₁NH, br, s, 1H), 9.76(C₅NH, s, 1H), 9.19 (C₅NH and C₄NH, s, 2H), 8.74 (C₉NH, m, 1H), 7.83(C₃₅NH, br, s, 3H, protonated ammonium), 4.41-4.32 (C₆H, m, 2H), 3.98(C₃₁H, br, s, 1H), 3.29 (C₇H, br, m, 2H), 2.94 (C₉H, br, m, 3H), 2.74(C₃₅H, br, m, 2H), 1.86 (C₃₂H, m, 1H), 1.77 (C₃₂H, m, 1H), 1.53 (C₃₄H,m, 2H), 1.43 (C₃₃H, m, 1H), 1.29 (C₃₃H, m, 1H). ¹³C-NMR (125 MHz, D₂Oand a drop of TFA) δ (ppm): 171.08 (C₃₀), 162.60 (TFA), 160.39 (C₄),155.87 (C₅), 155.73 (C₂₀), 149.77 (C₂), 118.39 (C₁), 114.54 (CF₃ fromTFA), 82.94 (C₃), 60.04 (C₃₁), 44.33 (C₆), 39.17 (C₇), 38.90 (C₃₅),28.03 (C₃₄), 26.44 (C₃₂), 26.37 (C₃₃), 21.52 (C₉). Positive ESI-MS:Expected mass for (M+H⁺)/z, 381.19. Observed, 381.20 ((M+H⁺)/z, 100%).Negative ESI-MS: Expected mass for (M−H⁺)/z, 379.19. Observed, 379.30((M−H⁺)/z, 100%). High resolution ESI-MS: Expected mass for (M+H⁺)/z,381.1999. Observed, 381.2003. Elemental analysis: Calculated for[C₁₅H₂₄N₈O₄—(CF₃CO₂H)_(2.5)—(H₂O)_(1.5)]: C, 34.69; H, 4.2, N, 16.18; F:20.58. Found: C, 34.99; H, 4.16; N, 15.96; F, 20.26.

A 94% solution of TFA/thioanisole (15 mL) was added to D-16 (0.93 g,0.93 mMol) at rt under N₂ atmosphere. After stirring for 86 h at rt,Et₂O (50 mL) was added. The precipitate formed was centrifuged andwashed with Et₂O (3×10 mL), redissolved in TFA (15 mL), reprecipitatedwith Et₂O (20 mL) then washed with Et₂O (2×10 mL) and CH₃OH (10 mL).D-Module 1 was obtained as a white solid(C₁₅H₂₄N₈O₄—(CF₃CO₂H)_(2.5)—(H₂O)_(1.5), 0.36 g, 50%). mp>298° C.(decomposed). ¹H-NMR (DMSO-d₆, 500 MHz) δ (ppm): 12.56 (C₃₀OH or C₃₁NH,br, s, 1H), 9.48 (C₅NH, s, 1H), 9.23 (C₅NH and C₄NH, s, 2H), 8.61 (C₉NH,m, 1H), 7.79 (C₃₅NH, br, s, 3H, protonated ammonium), 4.41-4.32 (C₆H, m,2H), 3.99 (C₃₁H, br, s, 1H), 3.31 (C₇H, br, m, 2H), 2.95 (C₉H, br, m,3H), 2.76 (C₃₅H, br, m, 2H), 1.86 (C₃₂H, m, 1H), 1.77 (C₃₂H, m, 1H),1.54 (C₃₄H, m, 2H), 1.43 (C₃₃H, m, 1H), 1.30 (C₃₃H, m, 1H). ¹³C-NMR (125MHz, D₂O) δ (ppm): 172.19 (C₃₀), 162.47 (TFA), 160.2 (C₄); 155.67 (C₅),155.46 (C₂₀), 149.69 (C₂), 117.42 (C₁), 115.09 (CF₃ from TFA), 82.74(C₃), 61.32 (C₃₁), 44.25 (C₆), 38.89 (C₇), 38.68 (C₃₅), 28.51 (C₃₄),27.62 (C₃₂), 26.37 (C₃₃), 21.32 (C₉). Positive ESI-MS: Expected mass for(M+H⁺)/z, 381.19. Observed, 381.1 ((M+H⁺)/z, 100%).

A 94% solution of TFA/thioanisole (7.5 mL) was added to 18 (0.4 g, 0.48mMol) at rt under N₂ atmosphere. After stirring for 4 h at rt, Et₂O (50mL) was added. The precipitate formed was centrifuged and washed withEt₂O (3×10 mL), redissolved in TFA (15 mL), reprecipitated with Et₂O (20mL) then washed with Et₂O (2×10 mL) and CH₃OH (10 mL). Module 4 wasobtained as a white solid (C₁₄H₂₄N₈O₄—(CF₃CO₂H)₂—(H₂O)_(1.5), 0.21 g,77%). mp=130° C. ¹H-NMR (500 MHz, 90% H₂O/D₂O) δ (ppm): 9.14 (C₅NH, br,s, 2H,), 8.32 (C₄NH, br, m, 1H), 7.53-7.29 (C₇NH, C₉NH, C₃₅NH, m, 4H),3.30 (C₇H, s, 2H), 2.93-2.83 (C₉H, C₃₁H, C₃₅H, m, 7H), 1.55-1.50 (C₃₂,C₃₄H, m, 4H), 1.28 (C₃₃H, t, J=4.8 Hz, 2H). C₆H suppressed with thesolvent. ¹³C-NMR (75 MHz, D₂O) δ (ppm): 163.6-163.2 (TFA), 160.42 (C₄),155.92 (C₂₀), 155.74 (C₅), 149.89 (C₂), 118.48 (C₁), 114.61 (CF₃ fromTFA), 82.92 (C₃), 47.92 (C₆), 45.80 (C₇), 39.31 (C₃₁, C₃₅), 27.98 (C₃₂),26.41 (C₃₃), 25.16 (C₃₄), 22.92 (C₉). Positive ESI-MS: Expected mass for(M+H⁺)/z, 337.20. Observed 337.2 ((M+H⁺)/z, 100%). Negative ESI-MS:Expected mass for (M−H⁺)/z, 335.20. Observed 335.3 ((M−H⁺)/z, 20%).

3-bromo-1-propanol (10 g, 71.9 mMol) was cooled to 0° C. in asingle-neck round-bottom flask (100 mL), and DHP (7.22 mL, 79.1 mMol)was added dropwise. 12 M HCl (2 drops) was added, and the reactionmixture was stirred at rt for 2 h. The reaction was quenched with Na₂CO₃(1.0 g), and the solid was filtered and washed with CH₂Cl₂ (20 mL). Thefiltrate was evaporated under reduced pressure (rotavap), and theresidual oil was purified by low-pressure short path distillation (0.05mm Hg). Compound 19 (C₈H₁₅BrO₂, 15.0 g, 94%) was obtained as a colorlessoil. R_(f)=0.45 (10% EA/Hex). ¹H NMR (CDCl₃, 200 MHz) δ (ppm): 4.60(C₄H, t, J=3.4 Hz, 1H), 3.84 (C₈H, m, 2H), 3.43-3.54 (C₁H, C₃H m, 4H),2.10 (C₂H, m, 2H), 1.49-2.0 (C₅H-C₇H, m, 6H). ¹³C NMR (CDCl₃, 50 MHz) δ(ppm): 98.78 (C₄), 64.77 (C₈), 62.14 (C₃), 32.82, 30.58, 30.50, 25.33,19.39 (C₁, C₂, C₅-C₇).

Malononitrile (8.88 g, 135 mMol) in DME (63 mL) was added dropwise to asuspension of 95% NaH (1.93 g, 80.4 mMol) and DME (30 mL) in athree-neck round-bottom flask (500 mL). Compound 19 (15.0 g, 67.0 mMol)in DME (14 mL) was added dropwise, and the reaction mixture was stirredat rt overnight under N₂ atmosphere. The reaction was quenched with 5%aqueous NH₄Cl until pH ˜8. The reaction mixture was diluted with EA (100mL) and washed with dH₂O (3×200 mL) and brine (100 mL). The organiclayer was dried over anhydrous Na₂SO₄, filtered, and the solvents wereevaporated under reduced pressure (rotavap). Compound 20 (C₁₁H₁₆N₂O₂,12.2 g, 88%) was obtained as a colorless oil after silica gel flashchromatography. (0-15% Et₂O/Hex). R_(f)=0.35 (70% Et₂O/Hex). ¹H NMR(CDCl₃, 200 MHz) δ (ppm): 5.00 (C₄H, t, J=2.8 Hz, 1H), 4.00 (C₉H, t,J=7.2 Hz, 2H), 3.77 (C₈H, m, 2H), 3.45 (C₃H, p, J=5.3 Hz, 2H), 2.12(C₁H, q, J=7.5 Hz, 2H), 1.46-1.90 (C₅H-C₇H, C₂H m, 8H). ¹³C NMR (CDCl₃,50 MHz) δ (ppm): 112.77 (C₁₀, C₁₁), 98.86 (C₄), 65.56 (C₈), 62.43 (C₃),30.34, 28.48, 26.04, 25.03, 22.10, 19.44 (C₁, C₂, C₅-C₇, C₉). CI-MS:Expected mass for (M+H⁺)/z, 209.12. Observed, 209 ((M+H⁺)/z, 67%), 125((M+H⁺)/z —CH₂CH₂CH₂OTHP, 10%). Positive high resolution CI-MS: Expectedmass for (M+H⁺)/z, 208.1212. Observed, 208.1217.

EtOH (250 mL) and sodium (2.71 g, 118 mMol) were stirred in a two-neckround-bottom flask (500 mL) under N₂ atmosphere until dissolution of thesodium. The solution was transferred to compound 20 (12.2 g, 58.6 mMol)and guanidinium hydrochloride (6.71 g, 70.3 mMol) in a three-neckround-bottom flask (500 mL), and the reaction mixture was refluxed for 5h under N₂ atmosphere. The reaction was then cooled to rt and quenchedwith 1M aqueous HCl until pH ˜8. The solvents were evaporated underreduced pressure (rotavap) to yield compound 21 (C₁₂H₂₁N₅O₂, 9.15 g,74%) as a light yellow solid after silica gel flash chromatography(0-10% MeOH/CH₂Cl₂). R_(f)=0.08 (10% MeOH/CH₂Cl₂). mp=170° C. ¹H NMR(DMSO-d-₆, 200 MHz) δ (ppm): 7.17 (C₁₂NH, s, 2H), 7.04 (C₁₀NH, C₁₁NH, s,4H), 4.52 (C₄H, m, 1H), 3.70 (C₈H, m, 2H), 3.46 (C₃H, m, 2H), 2.28 (C₁H,m, 2H), 1.45-1.73 (C₅H-C₇H, C₂H, m, 8H). ¹³C NMR (DMSO-d₆, 50 MHz) δ(ppm): 208.04 (C₁₀, C₁₁), 152.41 (C₁₂), 99.34, 83.58 (C₄, C₉), 61.66,66.20 (C₃, C₈), 56.05, 30.46, 27.42, 25.07, 19.43 (C₁, C₂, C₅-C₇).CI-MS: Expected mass for (M+H⁺)/z, 268.17. Observed, 268 ((M+H⁺)/z,100%), 184 ((M+H⁺)/z-THP, 28%). Positive high resolution FAB-MS (KIPEG,NBA): Expected mass for (M+H⁺)/z, 268.1774. Observed, 268.1786.

Compound 21 (2.0 g, 7.44 mMol), THF (200 mL), Boc₂O (11.43 g, 52.4mMol), Et₃N (8.34 mL, 59.9 mMol), and DMAP (0.64 g, 5.24 mMol) werestirred in a single-neck round-bottom flask (500 mL) overnight at rtunder N₂ atmosphere. The reaction was quenched with 5% aqueous NaHCO₃ (5mL), and the THF was evaporated under reduced pressure (rotavap). Theresulting material was dissolved in Et₂O (100 mL), and the organic layerwas washed with 5% aqueous NaHCO₃ (2×100 mL), dH₂O (100 mL) and brine(50 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered,and the Et₂O was evaporated under reduced pressure (rotavap). Compound22 (C₄₂H₆₉N₅O₁₄, 2.9 g, 45%) was obtained as a foam after silica gelflash chromatography (0-15% EA/Hex). R_(f)=0.29 (20% EA/Hex). mp=104° C.¹H NMR (CDCl₃, 200 MHz) δ (ppm): 4.58 (C₄H m, 1H), 3.74 (C₈H, m, 2H),3.44 (C₃H m, 1H), 2.58 (C₁H, m, 2H), 1.10-1.90 (C₁₅H, C₁₈H, C₂₁H, C₂₄H,C₅H-C₇H, C₂H, m, 62H). ¹³C NMR (CDCl₃, 125 MHz) δ (ppm): 161.28 (C₁₀,C₁₁), 155.45 (C₁₂), 150.09 (C₁₃, C₁₈), 149.48 (C₁₉, C₂₂), 127.66 (C₉),97.88 (C₄), 83.38 (C₂₀, C₂₃), 82.77 (C₁₄, C₁₇), 65.89, 61.47 (C₃, C₈),27.30 (C₂₁, C₂₄), 27.25 (C₁₅, C₁₈), 30.16, 25.03, 23.07, 22.13, 18.86(C₁, C₂, C₅-C₇). CI-MS: Expected mass for (M+H⁺)/z, 868.49. Observed,868 ((M+H⁺)/z, 4%), 784 ((M+H⁺)/z-THP, 3%), 768 ((M+H⁺)/z-Boc, 9%), 694((M+H⁺)/z-2Boc-THP, 20%.). Positive high resolution FAB-MS (KIPEG, NBA):Expected mass for (M+H⁺)/z, 868.4919. Observed, 868.4903.

Compound 22 (2.8 g, 3.22 mMol), MeOH (100 mL) and pTsOH (0.06 g, 0.32mMol) were stirred in a single-neck round-bottom flask (250 mL) for 5 hunder N₂ atmosphere. The reaction was quenched with 5% aqueous NaHCO₃ (3mL), and the solvents were evaporated under reduced pressure (rotavap).The solid material was dissolved in Et₂O (100 mL) and washed with 5%aqueous NaHCO₃ (5×100 mL), dH₂O (100 mL), and brine (50 mL). The organiclayer was dried over anhydrous Na₂SO₄, filtered, and the solvent wasevaporated under reduced pressure (rotavap) to give compound 23(C₃₇H₆₁N₅O₁₃, 2.47 g, 98%) as a foam. R_(f)=0.13 (30% EA/Hex). mp=148°C. ¹H NMR (CDCl₃, 200 MHz) δ (ppm): 3.55 (C₃H, m, 2H), 2.59 (C₁H, t,J=7.8 Hz, 2H), 1.81 (C₂H, m, 2H), 1.5-1.6 (C₁₅H, C₁₈H, C₂₁H, C₂₄H, m,54H). ¹³C NMR (CDCl₃, 50 MHz) δ (ppm): 161.39 (C₁₀, C₁₁), 155.57 (C₁₂),150.17 (C₁₃, C₁₆), 149.98 (C₁₉, C₂₂), 127.49 (C₉), 83.83 (C₂₀, C₂₃),83.05 (C₁₄, C₁₇), 61.22 (C₃), 29.98 (C₂), 27.44 (C₁₅, C₁₈, C₂₁, C₂₄),22.54 (C₁). CI-MS: Expected mass for (M+H⁺)/z, 783.43. Observed, 783((M+H⁺)/z), 684 ((M+H⁺)/z-Boc): Positive high resolution FAB-MS (KIPEG,NBA): Expected mass for (M+H⁺)/z, 784.4344. Observed, 784.4323.

Compound 23 (0.20 g, 0.26 mMol), CH₂Cl₂ (10 mL) and Dess-Martinperiodinane (0.22 g, 0.51 mMol) (Dess J. Org. Chem. 1983, 48, 4155-4156,incorporated herein by refernce.) were stirred in a single-neckround-bottom flask (50 mL) at rt for 2 h. The reaction was quenched with5% aqueous NaHCO₃ (2 mL), diluted with CH₂Cl₂ (50 mL) and washed with 5%aqueous NaHCO₃ (10×50 mL) and brine (50 mL). The organic layer was driedover anhydrous Na₂SO₄, filtered, and the solvent was evaporated underreduced pressure (rotapvap) to give compound 24 (C₃₇H₅₉N₅O₁₃, 0.19 g,94%) as a foam. R_(f)=0.34 (30% EA/Hex). mp=98-99° C. ¹H NMR (CDCl₃, 200MHz) δ (ppm): 9.77 (C₃H, s, 1H), 2.77 (C₁H, m, 2H), 1.2-1.28 (58H, C₁₅H,C₁₇H, C₂₁H, C₂₄H, C₂H, m). ¹³C NMR (CDCl₃, 50 MHz) δ (ppm): 200.00 (C₃),162.11 (C₁₀, C₁₁), 156.77 (C₁₂), 150.91 (C₁₃, C₁₆), 150.33 (C₁₉, C₂₂),127.19 (C₉), 84.62 (C₂₀, C₂₃), 83.94 (C_(14,) C₁₇), 41.93 (C₂), 28.14(C₂₁, C₂₄), 27.93 (C₁₅, C₁₈), 19.05 (C₁). CI-MS: Expected mass for(M+H⁺)/z, 782.42. Observed, 782 ((M+H⁺)/z, 3%), 754 ((M+H⁺)/z-CO, 0.4%),682 ((M+H⁺)/z-Boc, 11%), 582 ((M+H⁺)/z-2Boc, 4%). Positive highresolution FAB-MS (KIPEG, NBA): Expected mass for (M+H⁺)/z, 782.4188.Observed, 782.4171.

Compound 24 (0.36 g, 0.455 mmol) and N(ACO)₃BH (0.131 g, 0.621 mMol)were added in one portion to a vigorously stirred solution of L-15 (0.20g, 0.41 mmol), 1,2-DCE (3 mL) and Et₃N (0.17 mL, 1.24 mmol) in a singleneck round-bottom flask (50 mL). The reaction mixture was stirredovernight at rt under N₂ atmosphere then quenched with 5% aqueous NaHCO₃(2 mL). CH₂Cl₂ (25 mL) was added, and the organic layer was separatedand washed with 5% aqueous NaHCO₃ (2×25 mL), dH₂O (25 mL) and brine (26mL), dried over anhydrous Na₂SO₄, filtered, and evaporated under reducedpressure (rotavap). Compound 25 (C₅₉H₉l N₇O₁₆Si, 0.22 g, 46%) wasobtained as a foam after silica gel flash chromatography (0-20% EA/Hex).R_(f)=0.2 (30% EA/Hex). mp=128° C. (decomposition). ¹H NMR (CDCl₃, 500MHz) δ (ppm): 7.27-7.39 (C₃₉H-C₄₃H, m 5H), 5.23 (C₄₁NH, br, s, 1H), 5.08(C₄₃H, s, 2H), 4.18 (C₃₁H, t, J=8 Hz, 2H), 3.20 (C₃₆H, m, 2H), 3.10(C₃₈H, t, J=6.5 Hz, 1H), 2.30-2.55 (C₁H, C₃H, m, 4H), 1.37-1.48 (C₁₅H,C₁₈H, C₂₄H, C₃₈H-C₄₀H, C₂H, m, 62H), 0.98 (C₂₈H, t, J=8.5Hz, 2H), 0.00(C₂₇H, s, 9H). ¹³C NMR (CDCl₃, 125 MHz) δ (ppm): 175.96 (C₃₀), 161.86(C₁₀, C₁₁), 156.72, (C₁₂, C₃₆), 150.77 (C₁₃, C₁₆), 150.17 (C₁₉, C₂₂),130.01 (C₃₈), 128.42 (C₉), 128.13 (C₄₀, C₄₂), 128.63 (C₃₉, C₄₃), 128.21(C₄₁), 84.15 (C₂₀, C₂₃), 83.50 (C₁₄, C₁₇), 66.57, 63.05 (C₃₇, C₂₉),61.42 (C₃₁), 47.67 (C₃), 40.96 (C₃₅), 33.61 (C₃₂), 29.97, 28.21 (C₂,C₃₄), 28.00, 27.95 (C₁₅, C₁₈, C₂₁, C₂₄), 24.37, 23.44 (C₁, C₃₃) 17.71(C₂₈), −1.31 (C₂₇). ESI-MS: Expected mass for (M+H⁺)/z 1146.63.Observed, 1146.47.

Compound 25 (0.20 g, 0.18 mMol) and 94% TFA/thioanisole (10 mL) werestirred in a single neck round-bottom flask (50 mL) overnight at rtunder N₂ atmosphere. The TFA was evaporated and the crude product wassuspended in Et₂O (20 mL), centrifuged down (5000 rpm, 10 min), and thesolid isolated was resuspended in Et₂O (9 mL). Thissuspension/centrifugation/isolation procedure was repeated three times.After drying under vacuum, L-module 2(C₁₃H₂₅N₇O₂—(CF₃CO₂H)_(2.5)—(H₂O)_(2.0)—(Et₂O)_(0.5), 0.10 g, 82%) wasobtained as an off-white hygroscopic solid. mp=108° C. ¹H NMR (DMSO-d₆,500 MHz) δ (ppm): 8.85-9.10 (C₃₀OH, br, 1H,), 7.70-7.90 (C₃₅NH, br, 3H),6.90-7.20 (C₁₀NH-C₁₂NH, br, 6H), 3.91 (C₃₁ H, br, 1H), 2.95 (C₃₅H, br,2H), 2.76 (C₃H, br m, 2H), 2.31 (C₁H, br m, 2H), 1.30-1.85 (C₃₂H-C₃₄H,C₂H, br m, 8H). ¹³C-NMR (DMSO-d₆, 75 MHz) δ (ppm): 170.68 (C₃₀), 158.01(C₁₁), 153.08 (C₁₂), 83.39 (C₉), 59.65 (C₃₁), 46.06 (C₃), 29.00, 27.07(C₂, C₃₄), 24.35 (C₃₂), 21.99 (C₃₃), 19.59 (C₁). CI-MS: Expected massfor (M+H⁺)/z, 312.21. Observed, 312 ((M+H⁺)/z, 5%), 294 ((M+H⁺)/z-H₂O,100%). Elemental analysis: Calculated for[C₁₃H₂₅N₇O₂—(CF₃CO₂H)_(2.5)—(H₂O)_(2.0)—(Et₂O)_(0.5)]: C, 35.82; H,5.26; N, 15.13; F, 21.68. Found: C, 36.12; H, 5.07; N, 14.86; F, 21.55.

EtOH (12 mL) and sodium (0.187 g, 8.12 mMol) were stirred under N₂atmosphere in a two-neck round-bottom flask (50 mL) until dissolution ofthe sodium. Diethyl malonate (0.95 mL, 6.24 mMol) was added to thesolution, and the reaction mixture was stirred at rt for 5 min. Aftercooling to −50° C., allyl bromide (0.82 g, 6.87 mMol) was addeddropwise. The reaction mixture was then allowed to warm to rt and wasstirred overnight. The reaction was quenched with 5% aqueous NH₄Cl untilneutral pH, the solvents were evaporated under reduced pressure(rotavap), and the crude product was dissolved in CH₂Cl₂ (20 mL) andwashed with dH₂O (2×20 mL). The organic layer was dried over anhydrousNa₂SO₄, filtered, and the solvent was evaporated under reduced pressure(rotavap). Compound 26 (C₁₀H₁₆O₄, 0.92 g, 73%) was obtained as acolorless oil after silica gel flash chromatography (0-1.5% EA/Hex).R_(f)=0.36 (10% EA/Hex). ¹H NMR (CDCl₃, 500 MHz) δ (ppm): 5.81-5.88(C₉H, m, 1H), 5.0-5.88 (C₁₀H m, 2H), 4.19 (C₂H, C₆H, q, J=3.4 Hz, 12 Hz,4H), 2.64 (C₄H, t, J=6.9 Hz, 1H), 1.26 (C₇H, t, J=9.56 Hz, 6H).

EtOH (1 L) and sodium (12.83 g, 558 mMol) were stirred in a single-neckround-bottom flask (2 L) under N₂ until dissolution of the sodium. Tothis solution urea (27.9 g, 465 mMol) and compound 27 (93.0 g, 465 mMol)were added, and the mixture was refluxed for 5 h. After cooling to rt,the reaction was quenched with 1M HCl until neutral pH. The precipitateformed was filtered and washed with EtOH (50 mL) and Et₂O (50 mL), anddried under vacuum to yield compound 27 (C₉H₁₀N₂O₃, 52.0 g, 67%) as awhite solid. ¹H NMR (DMSO-d₆, 200 MHz) δ (ppm): 9.41 (C₁₁NH, s, 2H),6.26-6.30 (C₉H, m, 1H), 5.06-5.33 (C₁₀H, m, 2H), 3.21 (C₈H, d, J=8.0 Hz,2H).

A solution of compound 27 (47.0 g, 280 mmol) and POCl₃ (208 mL, 223mMol) was heated to 70° C. in a single neck round-bottom flask (500 mL),and N,N-dimethyl aniline (9.68 g, 76.5 mMol) was added dropwise. Themixture was refluxed for 4 h under N₂ atmosphere, then cooled to rt.Excess POCl₃ was removed under reduced pressure (rotavap), and theslurry was slowly poured on crashed ice (500 g). The resulting aqueousmixture was extracted with CH₂Cl₂ (3×100 mL), the organic layers werecombined and washed with brine (100 mL), dried over anhydrous Na₂SO₄,filtered, and evaporated under reduced pressure (rotavap). Compound 28(C₇H₅C₁₃N₂, 44 g, 71%) was obtained as a crystalline solid after silicagel flash chromatography (0-2% Et₂O/Hex). R_(f)=0.46 (10% EA/Hex). ¹HNMR (CDCl₃, 200 MHz) δ (ppm): 5.67-5.89 (C₉H, m, 1H), 5.0-5.2 (C₁₀H, m,2H), 3.58 (C₈H, d, J=6.2 Hz, 2H).

Benzyl alcohol (0.56 mL, 5.40 mMol) was added dropwise to a suspensionof 95% NaH (0.134 g, 5.58 mMol) and THF (18 mL) in a two-neckround-bottom flask (1 L) under N₂ atmosphere. After cooling to 0° C.(ice bath) compound 28 (0.20 g, 0.9 mMol) was added in one portion, andthe reaction mixture was refluxed overnight. The reaction was thenquenched with 5% aqueous NH₄Cl until neutral pH. After adding brine (200mL), the mixture was extracted with EA (2×20 mL), the organic layerswere combined, dried over anhydrous Na₂SO₄, filtered, and the solventwas evaporated under reduced pressure (rotavap). Compound 29(C₂₈H₂₆N₂O₃, 0.30 g, 77%) was obtained as a white solid after silica gelflash chromatography (0-1.5% EA/Hex). R_(f)=0.50 (15% EA/Hex). mp=76-77°C. ¹H NMR (CDCl₃, 200 MHz) δ (ppm): 7.21-7.44 (CH₁₄-C₁₈H, C₂₁H-C₂₅H, m,15H), 5.74-5.95 (C₉H, m, 1H), 5.41 (C₁₂H, C₁₉H, br, J=4.6 Hz, 6H),4.90-5.05 (C₁₀H, m, 2H), 3.33 (C₈H, d, 2H, J=3.3 Hz). ¹³C NMR (CDCl₃, 50MHz) δ (ppm): 169.54 (C₃, C₅), 162.52 (C₁₁), 137.51, 135.96 (C₉, C₁₃,C₂₀), 128.95-127.98 (C₁₄-C₁₈, C₂₁-C₂₅), 115.45 (C₁₀), 96.86 (C₄), 69.52(C₁₂), 69.18 (C₁₉), 27.42 (C₈). CI-MS: Expected mass for (M+H⁺)/z,439.20. Observed, 439 ((M+H⁺)/z, 100%). Positive high resolution FAB-MS(NBA, KIPEG): Expected mass for (M+H⁺)/z, 439.2022. Observed, 439.2027.

Compound 29 (10.0 g, 22.83 mMol), dioxane (200 mL), dH₂O (50 mL), NaIO₄(14.59 g, 1.14 mMol), OsO₄ (0.1 M in t-BuOH, 2.23 mL, 0.114 mMol) werestirred in a single neck round-bottom flask (500 mL) overnight under N₂atmosphere. The reaction was quenched with a 3% aqueous solution ofsodium sulfite until the solution turned colorless. The off-whiteprecipitate formed was dissolved in EA (200 mL) and washed with dH₂O(3×200 mL) and brine (100 mL). The organic layer was separated, driedover anhydrous Na₂SO₄, filtered, and the solvent was evaporated underreduced pressure (rotavap). Compound 30 (C₂₇H₂₄N₂O₄, 7.30 g, 73%), wasobtained as a white solid after silica gel flash chromatography (5-25%EA/Hex). R_(f)=0.31 (15% EA/Hex). mp=81-82° C. ¹H NMR (CDCl₃, 200 MHz) δ(ppm): 9.63 (C₉H, s, 1H), 7.33-7.52 (C₁₄H-C₁₈H, C₂₁H-C₂₅H, m, 15H), 5.43(C₁₂H, C₂₀H, s, 6H), 3.62 (C₈H, s, 2H). ¹³C NMR (CDCl₃, 75 MHz) δ (ppm):198.81 (C₉), 169.27 (C₃, C₅), 136.80 (C₁₂), 136.50 (C₁₉), 128.28 (C₁₅,C₁₇, C₂₂, C₂₄), 127.84 (C₁₆, C₂₃), 127.40 (C₁₄, C₁₈, C₂₁, C₂₅), 89.49(C₄), 69.21 (C₁₂), 68.77 (C₁₉), 37.03 (C₈). CI-MS: Expected mass for(M+H⁺)/z, 441.18. Observed, 441 ((M+H⁺)/z, 100%). Positive highresolution FAB-MS (NBA, KIPEG): Expected mass for (M+H⁺)/z, 441.1820.Observed, 441.1814.

Compound 30 (0.26 g, 0.60 mMol) and Na(AcO)₃BH (0.19 g, 0.91 mMol) wereadded in one portion to a vigorously stirred solution of L-15 (0.30 g,0.60 mMol), 1,2-DCE (5 mL) and Et₃N (0.12 mL, 0.65 mMol) in a singleneck round-bottom flask (100 mL). The reaction mixture was stirredovernight at rt under N₂ atmosphere then quenched with 5% aqueous NaHCO₃(2 mL). CH₂Cl₂ (30 mL) was added and the organic layer was separated andwashed with 5% aqueous NaHCO₃ (3×30 mL) and brine (30 mL), dried overanhydrous Na₂SO₄, filtered, and evaporated under reduced pressure(rotavap). Compound 31 (C₄₆H₅₆N₄O₇Si, 0.38 g, 55%) was obtained as afoam after silica gel flash chromatography (5-15% EA/Hex). R_(f)=0.44(30% EA/Hex). ¹H NMR (CDCl₃, 200 MHz) δ (ppm): 7.24-7.51 (C₂₁H-C₂₅H,C₁₄H-C₁₈H, C₃₉H-C₄₃H, m, 20H), 5.38 (C₁₉H, s, 4H,), 5.13 (C₁₂H, s, 2H),5.11 (C₃₇H, s, 2H), 4.95 (C₃₅NH, br, m, 1H), 4.17 (C₂₉H, t, J=8.6 Hz,2H), 3.40-3.00 (C₃₁H, C₃₅H, m, 3H), 2.90-2.60 (C₈H, C₉H, m, 4H),1.90-1.20 (C₃₂H-C₃₄H, m, 6H), 0.97 (C₂₈H, t, J=8.6 Hz, 2H), 0.07 (C₂₇H,s, 9H). ¹³C NMR (CDCl₃, 50 MHz) δ (ppm): 175.25 (C₃₀) 169.19 (C₃, C₅),161.73 (C₁₁), 156.26 (C₃₆), 136.87, 136.78, 136.59 (C₃₈, C₁₃, C₂₀),128.35, 128.28, 127.95, 127.88, 127.81, 127.72, 127.40, 126.96 (C₁₄-C₁₈,C₂₁-C₂₅, C₃₉-C₄₃), 95.73 (C₄), 68.83 (C₁₂), 68.09 (C₁₉), 66.34 (C₃₇),62.68 (C₂₉), 61.05 (C₃₁), 46.71 (C₉), 40.68 (C₃₅), 32.72 (C₃₂), 29.51(C₃₄), 22.72 (C₈, C₃₃), 17.31(C₂₈), −1.50 (C₂₇). PD-MS: Expected massfor (M+H⁺)/z, 805.40. Observed, 805. Positive high resolution FAB-MS(NBA, KIPEG): Expected mass for (M+H⁺)/z, 805.3997. Observed, 805.3984.

Compound 31 (0.25 g, 0.31 mMol) and 94% TFA/thioanisole (10 mL) werestirred in a single neck round-bottom flask (50 mL) for 96 h at rt underN₂ atmosphere. The TFA was evaporated, and the crude product wassuspended in Et₂O (20 mL), centrifuged down (5000 rpm, 10 min), and thesolid isolated was resuspended in Et₂O (9 mL). Thissuspension/centrifugation/isolation procedure was repeated three times.After drying under vacuum, L-module 3(C₁₂H₂₀N₄O₅—(CF₃CO₂H)₂—(H₂O)_(1.0)—(Et₂O)_(0.25)], 0.11 g, 65%) wasobtained as an off-white hygroscopic solid. ¹H NMR (CDCl₃, 200 MHz) δ(ppm): 9.69 (C₃NH, C₅NH br s, 2H), 7.75 (C₃₅NH, br s, 3H), 2.75-3.11(C₉H, C₃₅H, m, 4H), 2.52 (C₈H, m, 2H), 1.31-1.97 (C₃₂H-C₃₄H, C₃₈H, m,6H). ¹³C NMR (CDCl₃, 75 MHz) δ (ppm): 171.76 (C₃₀) 166.34 (C₃, C₅),152.15 (C₁₁), 100.23 (C₄), 59.68 (C₇), 48.43 (C₅), 32.54 (C₃₄), 29.38(C₃₂), 27.17 (C₈), 22.01 (C₃₃). PD-MS: Expected mass for (M+H⁺)/z,301.14. Observed, 301.9. Positive high resolution FAB-MS (NBA, KIPEG):Expected mass for (M+H⁺)/z, 301.1512. Observed, 301.1526. Elementalanalysis: Calculated for[C₁₂H₂₀N₄O₅—(CF₃CO₂H)₂—(H₂O)_(1.0)—(Et₂O)_(0.25)]: C, 36.14; H, 4.73; N,9.92. Found: C, 35.93; H, 4.38; N, 9.80.

Modules 2-4 were prepared to establish the role of the stackinginteractions, hydrophobic effect, and the chirality of the side chain inthe assembly of the tubular architectures. A shown in FIG. 5, a 600 MHzfb-NOESY 2D-¹H NMR spectrum of a 0.015 M solution of L-module 1 in 90%H₂O/D₂O at 20° C. and the imino/amino region of the spectrum (enlargedsection) showed the formation of a six-membered supermacrocyclestructure held through hydrogen bonds. In particular, L-module 1displayed nuclear Overhauser effects (NOE's) not only between H^(A) andH^(B) but also between H^(B) and H^(C)/H^(D). Since H^(B) andH^(C)/H^(D) are too far apart to display any intra-modular NOE's, theobservation of an NOE between them confirms the existence ofinter-modular H-bonds. This result is further diagnostic of theformation of the supermacrocycle structure since no other NOE's or iminoproton signals resulting from non-assembled modules or non-specificaggregates thereof were observed. The fb-NOESY (flip-back-noesy)experiment of L-module 1 was recorded in a Shigemi NMR tube on a VarianUnity Plus 600 MHz spectrometer. The States Haberkorn method was used inorder to obtain a phase sensitive spectrum. A mixing time of 400 ms wasused and 2×320 increments were collected with 3648 points. The data wasprocessed using a combination of shifted sinebell and Gaussian windowfunctions (VNMR 6.1B). Zero filling to 8k points in t₂ and 2k points int₁ was applied. A spline baseline correction was applied in theacquisition dimension. The Proton assignments are as follows: 9.15(H^(C)/H^(D), br, s, 2H), 8.34 (H^(B), br, s, 1H), 7.57 (H^(A), br, m,1H), 7.39 (H^(E), br, s, 3H, protonated ammonium), 3.81 (C₃₁H, t, J=5.0Hz, 1H), 3.36 (C₇H, t, J=6.0 Hz, 2H), 2.90 (C₉H, br, m, 3H), 2.85 (C₃₅H,br, m, 2H), 1.84-1.79 (C₃₂H, m, 2H), 1.55 (C₃₄H, br, s, 2H), 1.36 (C₃₃H,br, m, 1 H), 1.29 (C₃₃H, br, m, 1 H). C₆H was suppressed with thesolvent peak.

In agreement with the above described NMR spectrum, electrosprayinonization mass spectrometry (ESI-MS) (Finnigan MATLCQ) of a 1 mMsolution of L-module 1 in CH₃OH/H₂O (1/1) displayed all the peakscorresponding to the non-covalent intermediate species (1-mer to 6-mer)of the parent supermacrocycle as shown in FIG. 6. In particular, thepeaks at 381.4, 760.5, 1140.4, 1521.9, 1901.6 and 2282.2 correspond tothe non-covalent intermediate species of the self-assembledsupermacrocycle (1-mer to 6-mer). Because of their charged nature theassemblies were characterized without introducing additional ions. Underthe same conditions an equimolar aqueous mixture of the L-modules 2 and3 did not undergo self-assembly.

Correlation between the supramolecular chirality of foliate- andG-quadruplexes and circular dichroism (CD) activity has been drawn onthe basis of exciton coupling theory. The signed order of the CD excitoncouplet is governed by the relative helicity of the relevant twoelectric dipole transition moments, one from each stacked base. As shownin FIG. 7, the CD spectra (Jasco J-810) of L-module 1 and its D-isomer(open diamonds) in an aqueous buffered solution display a typicalcouplet centered at 286 nm, characteristic of stacked bases in a helicalenvironment, that vanishes at higher temperatures. (Note MES:2-(N-Morpholino) ethanesulfonic acid.) Under identical conditions,L-modules 2 and 3 or an equimolar mixture of both, as well as theachiral module 4 were CD-silent. Furthermore, the position of the CDcouplet maxima (280 and 292 nm) are not identical to their correspondingmaximum in the absorption spectrum (285 nm, see FIG. 8: λ¹ _(max)=236 nm(ε=21,500 M⁻¹ cm⁻¹); λ² _(max)=285 nm (ε=12,500 M⁻¹ cm⁻¹), whichindicates that the CD spectra are caused by chirality and not by lineardichroism of partially oriented samples. In the latter case, differentlypolarized absorption bands would appear as positive and negative bandsin a CD spectrum, with their maxima at the same position as in theabsorption spectrum. Note also that L-module 1 does not form gels orliquid crystalline phases in water. At higher concentration thismaterial tends to precipitate out of the solution as a white solid.

The hyperchromicity observed upon deoxyribonucleic acids (DNA)denaturation is a powerful indicator of its stability and tertiarystructure. In this regard, the supramolecular outcome of L-module 1behaves very much like DNA. The absorbance spectrum undergoes acooperative hyperchromic effect as the temperature increases (meltingtemperature T_(m) ^(285 nm)=50° C.). The same transition was recorded byvariable temperature CD (λ=292 nm). In particular, as shown in FIG. 9,the disappearance of the CD peaks at higher temperatures demonstratesthat the recorded cotton effect is the result of supramolecularchirality rather than intrinsic molecular chirality of L-module 1. Also,the observation of several isosbectic points (224, 246, 286, 303 nm) isan indication that the melting transition in water takes place betweentwo main stable species (nanotube

[supermacrocycle]#

monomeric modules). A plot of the elliptcity at 291 nm as a function oftemperature yields a cooperative melting transition around 50° C., inagreement with the transition recorded in the absorbance mode as shownin FIG. 10. In particular, FIG. 10 shows a Melting curve of 5×10⁻⁵ Msolution of L-module 1 in 0.01 M PIPES, pH 7.5. λ=285 nm, T_(ramp)=1°C./min. T_(m)=50° C. The wide transition temperature (from 35° to 70°C.) is presumably due to the length polydispersity of the sample.

In combination with NMR, ESI-MS, and CD it is clear that: (a) L-module 1undergoes a cooperative, hierarchical self-assembly process throughH-bonding, stacking interactions and hydrophobic effects and (b) theassembly generated displays supramolecular chirality, as a result ofhelically stacked supermacrocycles.

To assess the size of the assemblies generated, the dynamic lightscattering (DLS) spectrum was recorded for a buffered solution ofL-module 1. As specifically shown in FIG. 11, the dynamic lightscattering regularization diagram (Protein-Solutions MSTC200) of a 1.0mg/ml solution of L-module 1 in 0.01 M MES buffer, pH 5.5, 20° C. showedan average hydrodynamic radius of 30.4 nm (average of 15 measurments).The narrow distribution (92% in the range 19-69 nm) obtained with anaverage apparent hydrodynamic radius, R_(H), of 30.4 nm is rationalizedby invoking a columnar stack reminiscent of the solid or liquidcrystalline states of folate- and G-quadruplexes, and otherself-assembled materials. As control experiments, L-modules 2, 3, or anequimolar mixture of both did not generate any detectable aggregate.

Transmission electron micrograph (TEM) provided visual evidence of theformation of the nanotubular assemblies and confirmation of thespectroscopic data discussed above. FIG. 12 is a large area transmissionelectron micrograph of a negatively stained sample of L-module 1 (scalebare=100 nm). Formvar and carbon coated grids were floated on dropletsof 0.5 mg/ml in 0.01 M MES buffer at pH 5.5 for ˜30 s, transferred todroplets of 2% aqueous uranyl acetate for ˜30 s, blotted and viewed in aPhilips EM400 transmission electron microscope at 80 kV and 60,000×magnification. Images of the 8.75 nm lattice spacing of catalasecrystals, taken at the same magnification, were used for calibration.Negatives were scanned into a computer at 1600 dpi resolution and thediameter of nanotubes was determined using the image analysis program IPLab (Scanalytics, Fairfax, Va.). The measured outer diameter of ˜4.0 nm(this number includes a layer of the staining agent) is an average of200 measurements made on randomly selected nanotubes. In agreement withthe DLS data, most of the sample's length revolves around ˜60 nm.Although ˜10% of the sample forms bundles, there are no higher ordertwisted or helical aggregates, thereby suggesting that the CD spectrarecorded are the result of an intrinsic property of each individualnanotube. Since they are undetectable by DLS and because the sample wasfiltered on 0.2 μm filters prior to TEM imaging, the nanotube bundlesobserved arise most likely during solvent evaporation on the TEM grid.

It should be understood that the above described system establishes thatelectrostatic, stacking and hydrophobic interactions can be effectivelyorchestrated by hydrogen bonds to direct the hierarchical assembly andorganization of helical nanotubular architectures in an aqueous milieu.Furthermore, utilizing the above discussed synthetic strategy a widevariety of structurally different modules (i.e. molecules) can besynthesized. Including but not limited to, for example, a compoundhaving the formula:

wherein X is carbon or nitrogen; n is an integer of, 1, 2, 3, or 4; Y isan amino acid having an amino group covalently bound to an α-carbon ofsaid amino acid and said amino group is covalently bound to a carbon ofthe (CH₂)_(n) group; and R₁ is aliphatic; and salts thereof: a compoundhaving the formula:

wherein X is carbon or nitrogen; n is an integer of 1, 2, 3, or 4; R₁ isaliphatic; R₂ is hydrogen; and R₃ is —CH₃, —CH₂OH, —CH₂CH₂SCH₃,—CH₂CO₂H, —CH₂CH(CH₃)₂, —CH₂C₆H₅, —CH₂-p-(OH)C₆H₄, —CH₂CH₂CH₂CH₂NH₂, orR₂ and R₃ together form

and salts thereof: and a compound having the formula:

and salts thereof.

It should also be understood that the above described modules can beutilized in a method for forming nanotubes. In particular, the modulesof the present invention self assembly into nanotubes upon disposing thesame in an aqueous solution at an appropriate concentration, for examplein a pH 5.5 MES buffer at an appropriate concentration at 20° C.Furthermore, as previously mentioned it should be appreciated thathaving the ability to oligomerize and/or functionalize the modules atvirtually any position greatly enhances the ability to form a wide rangeof structurally unique modules and thus easily form a wide range ofnanotubes with specifically desired physical/chemical characteristics.The exact conditions for forming nanotubes with any one, or combinationof, the various modules of the present invention will depend upon theparticular characteristics of the module/modules utilized. However, inlight of the above disclosure these conditions are easily determinableby one of ordinary skill in the art from only routine experimentation.

While the invention has been illustrated and described in detail in thedrawings and foregoing description, such illustration and description isto be considered as exemplary and not restrictive in character, it beingunderstood that, only the preferred embodiments have been shown anddescribed and that all changes and modifications that come within thespirit of the invention are desired to be protected.

1. A compound having the formula:

wherein R₁is a halogen or

R₂ is a halogen or

R₃ is a halogen.
 2. The compound of claim 1 wherein: R₁ Cl, R₂ is Cl,and R₃ is Cl.
 3. The compound of claim 1 wherein: R₁ is

R₂ is Cl, and R₃ is Cl.
 4. The compound of claim 1, wherein: R₁ is

R₂ is

 and R₃ is Cl.
 5. A compound having the formula:

wherein R¹ is

and R₂ is


6. The compound of claim 5 wherein: R₂ is

and R₁ is


7. The compound of claim 5, wherein: R₁ is

and R₂ is


8. The compound of claim 5 wherein: R₁ is

and R₂ is


9. A compound having the formula:

wherein R₁ is

and R₂ is


10. The compound of claim 9 wherein: R₁ is

and R₂ is ^(—NH) ² .
 11. The compound of claim 9 wherein: R₁ is

and R₂ is


12. The compound of claim 9 wherein: R₁ is

and R₂ is


13. The compound of claim 9 wherein: R₁ is

and R₂ is


14. A compound having the formula:

wherein R¹ is


15. The compound of claim 14, wherein: R₁ is NH₂.
 16. The compound ofclaim 14, wherein: R₁ is